Caprazene as a novel compound and derivatives thereof, and caprazol as a novel compound and derivatives thereof

ABSTRACT

A caprazene compound is provided which is the compound represented by the following formula (Ia): 
     
       
         
         
             
             
         
       
     
     wherein Me stands for methyl group, and A is a hydrogen atom, and wherein said compound has the  1 H-NMR and  13 C-NMR data as set forth in Table 15.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a divisional of U.S. application Ser. No.12/345,190, filed Dec. 29, 2008, which is a divisional of U.S.application Ser. No. 10/543,887, filed Sep. 29, 2005, which is a 371application based on PCT/JP04/000969, filed Jan. 30, 2004, all of saidapplications incorporated herein by reference.

TECHNICAL FIELD

This invention relates to a novel compound, caprazene, which is preparedby acid hydrolysis of antibiotics, caprazamycins A, B, C, D, E, F and/orG in an aqueous solution of an acid, and also this invention relates toa novel compound, caprazol, which is prepared by hydrolysis ofcaprazamycins in an aqueous solution of an inorganic base. Caprazene andcaprazol are compounds represented by the stereo-structural formulae (I)and (IV) shown hereinafter, respectively, and they have no antibacterialactivity, but are useful as intermediate compounds usable for thesyntheses of a variety of antibacterial amide derivatives or esterderivatives therefrom. They are also useful as enzyme inhibitors havingan inhibitory activity against enzyme MraY which takes part in thebiosynthesis of the cell walls of bacteria.

This invention also includes a process for the preparation of caprazeneand a process for the preparation of caprazol, both of which comprise ahydrolysis of caprazamycins. This invention further relates to a varietyof novel caprazene amide derivatives or caprazene ester derivatives anda variety of novel caprazol amide derivatives or caprazol esterderivatives, all of which have antibacterial activities against variousbacteria. These antibacterial caprazene derivatives and caprazolderivatives according to this invention are expected to be useful forthe

therapeutic treatment of tuberculosis and treatment of bacterialinfections by atypical acid-fast bacteria, that is Mycobacterium AviumComplex (MAC) infection and other bacterial infections.

Further, this invention includes a variety of novel imidazolidinonederivatives (Code name: CP-IM) having the formula (VIII) hereinaftergiven which may be synthesized from caprazol through several reactionsteps and which have an antibacterial activity.

Furthermore, this invention relates to a pharmaceutical compositioncomprising as the active ingredient a caprazene derivative or a caprazolderivative or an imidazolidinone derivative as mentioned above.

Moreover, this invention includes novel uridine derivative of theformula (IX) hereinafter given which is prepared by treating caprazolwith methylamine to effect the ring-opening of the diazepine ring ofcaprazol and which is usable as an intermediate material for thesynthesis of the said imidazolidinone derivative CP-IM.

BACKGROUND ART

In the chemotherapy of bacterial infections, particularly thechemotherapy of infections by acid-fast bacteria, there have alreadybeen used, as antibacterial agent, antibiotics such as rifampicin,kanamycin, streptomycin, viomycin, capreomycin, cycloserine, and thelike.

A serious problem for the chemotherapy of the bacterial infections is inthat bacteria causative of the bacterial infections becomedrug-resistant. In particular, the appearance of acid-fast bacteriawhich are resistant to rifampicin, kanamycin, streptomycin, viomycin,capreomycin, cycloserine and the like has brought about a social problemin respect of the chemotherapy of the acid-fast bacterial infections.Thus, there is now a keen request for providing a novel chemotherapeuticagent which is effective against bacterial infections as induced by theacid-fast bacteria resistant to the antibacterial drugs. Stronglyrequested also is a novel chemotherapeutic drug that is effectiveagainst the bacterial infections which are induced by atypical acid-fastbacteria and for which no chemotherapeutic treatment has beenestablished yet.

In order to meet these requisites, therefore, there exists a strongdemand to find out or to create novel compounds which have novelchemical structures and can exhibit excellent properties such as highantibacterial activities in a different way from those of the knownantibiotics as hitherto utilized. We, the inventors of this invention,have carried out various investigations with the intention of providingnovel antibiotics having excellent antibacterial activities which canmeet the above-mentioned requisites.

Thus, there have already been proposed antibiotics, caprazamycins A, B,C, E and F which have been produced from Streptomyces sp. MK730-62F2strain (deposited under the depository number of FERM BP-7218 under theBudapest Treaty) and which exhibit high antibacterial activities againstacid-fast bacteria [see Pamphlet of PCT International Publication NumberWO 01/12643A1 and European patent application first publn. EP1211259A1].

Caprazamycins A, B, C, E and F are compounds represented by thefollowing general formula (A)

wherein R is tridecyl group for caprazamycin A, 11-methyl-dodecyl groupfor caprazamycin B, dodecyl group for caprazamycin C, undecyl group forcaprazamycin E, and 9-methyl-decyl group for caprazamycin F.

Further, there have also been provided caprazamycins D, G, D1 and G1which are produced from Streptomyces sp. MK730-62F2 strain (FERMBP-7218) (refer to the specification of PCT application No.PCT/JP02/13386 filed on Dec. 20, 2002).

Caprazamycin D and caprazamycin G are the compounds represented by thefollowing general formula (B)

wherein R is 10-methyl-undecyl group —(CH₂)₉CH(CH₃)₂ for caprazamycin Dand is 9-methyl-undecyl group —(CH₂)₈CH(CH₃)CH₂CH₃ for caprazamycin G.

Caprazamycin D1 and caprazamycin G1 are the compounds represented by thefollowing general formula (C)

wherein R is 10-methyl-undecyl group —(CH₂)₉CH(CH₃)₂ for caprazamycin D1and is 9-methyl-undecyl group —(CH₂)₈CH(CH₃)CH₂CH₃ for caprazamycin G1.

There have also been known liposidomycins A, B and C which are producedfrom Streptomyces glyceosporeus SN-1051M (FERM BP-5800) (refer toJapanese Patent Application First Publication KOKAI Sho-61-282088).

Liposidomycins A, B and C are compounds represented by the followinggeneral formula (D)

wherein R is 4,7-tridecadienyl group —(CH₂)₃—CH═CH—CH₂—CH═CH—(CH₂)₄—CH₃for liposidomycin A, 9-methyl-decyl group —(CH₂)₈—CH(CH₃)₂ forliposidomycin B and undecyl group —(CH₂)₁₀—CH₃ for liposidomycin C.

Further, there have been known liposidomycins G, H, K, L, M, N and Z andother liposidomycins' homologues (refer to Pamphlet of PCT InternationalPublication Number WO97/41248 and European Patent Application FirstPublication, EP 1001035A1).

Also known already are liposidomycins X-(III), Y-(III), Z-(III),C-(III), V-(III), A-(III), G-(III), M-(III), K-(III), and N-(III) (seeEuropean Patent Application First Publication EP 1001035A1) and they arecompounds represented by the following general formula (E)

wherein R is a long chain alkyl group shown in the Pamphlet ofWO97/41248 or in Table 1 of the European Patent Application PublicationEP1001035A1.

A report has been issued that relates to investigation on elucidation ofchemical structures of liposidomycins A, B and C [refer to The Journalof Organic Chemistry, Vol. 57, No. 24, pages 6392-6403 (1992)]. Thisreport describes (see the J.O.C., pages 6397-6399) three compounds,namely, compound 10 (given as anhydrodeacyl-liposidomycin havingmolecular weight of 557) of the following planar structural formula (F)

and Compound 11 (given as anhydrodeacyl-liposidomycin having molecularweight of 637) of the following planar structural formula (G)

and Compound 12 of the following planar structural formula (H)

each of the three compounds having been prepared by an alkalinehydrolysis of a mixture of liposidomycins B and C in a dilute aqueousNaOH solution at 37° C. to produce Compound 10 and Compound 11 and by areductive deacylation of a mixture of liposidomycins B and C with LiBH₄to produce Compound 12. The report also shows ¹³C-NMR data (Table III)and ¹H-NMR data (Table IV) of these three compounds, but thestereo-structures of these three compounds are unknown yet until now.

Caprazamycins A to G referred to in the above have one common skeletalstructure with each another and have excellent antibacterial activities.However, the antibacterial activities of caprazamycins A, B, C, D, E, Fand G are different, among them, depending upon the nature of bacteria.Further, upon the preparation of these caprazamycins by the cultivationof Streptomyces sp. MK730-62F2 strain referred to above as acaprazamycin-producing bacterial strain, followed by the recovery of thecaprazamycins from the resulting culture broth, it is usual that amixture of caprazamycins A to G is first obtained. Thus, it isnecessary, in order to separate the caprazamycins A to G from eachother, to carry out time-consuming and troublesome operationsnecessarily comprising the high performance liquid chromatography(HPLC).

Therefore, it has been requested to synthesize certain novelsemi-synthesized antibiotics which can have antibacterial activitiesequivalent or superior to those of caprazamycins A, B and C-G, and whichcan be prepared in an efficient way by utilizing a mixture comprisingtwo or more of caprazamycins A to G or by utilizing any one ofcaprazamycins A, B or C, alone. It has also been requested to providecertain novel semi-synthesized antibiotics which comprise the skeletalstructure common to caprazamycins A to G.

DISCLOSURE OF THE INVENTION

In order to satisfy the request as above-mentioned, we, the inventors ofthis invention, have made various investigations. First of all, we havecarried out some experiments wherein at least one of caprazamycins A toG, preferably caprazamycin B, is subjected to acid-hydrolysis in anaqueous acid solution, for example, an aqueous acetic acid solution at aconcentration of 50-90% by weight, or a dilute aqueous sulfuric acid orhydrogen chloride solution. As a result, we have found that theresulting reaction solution of the acid-hydrolysis of a caprazamycincontains the compound thus produced which is represented by thefollowing formula (I)

wherein Me stands for methyl group, and we have succeeded in isolatingsaid compound as a colorless solid. The compound of the formula (I)above has been recognized to comprise in its molecule a5′-substituted-uridine moiety and a 5-amino-5-deoxy-D-ribose moiety anda 1,4-diazepinone moiety having one double bond.

We have measured physicochemical properties and NMR data of the compoundso isolated. Further, the 5″-N-tert-butoxycarbonyl derivative has beenprepared from the said compound and then crystallized out. Thederivative obtained as the crystals has been analyzed by X-ray powderdiffractometry. Thus, the compound now isolated has been decided to havethe steric chemical structure shown by the formula (I) above.

Further, taking the physicochemical properties, ¹H-NMR data and ¹³C-NMRdata of said compound into consideration collectively, we have decidedthe said compound to be a novel substance and designated it ascaprazene.

By the way, when comparing the ¹³C-NMR data (Table III) and ¹H-NMR data(Table IV) of the Compound 10 of which stereo-structure is unknown yetand which is given by the planar structural formula on pages 6397-6399and page 6402 of the literature, The Journal of Organic Chemistry, Vol.57, No. 24 mentioned above with those of caprazene of the formula (I) ofthis invention, some of the data for the Compound 10 are not necessarilyconsistent with ¹³C-NMR data and ¹H-NMR data of caprazene of the formula(I) of this invention (refer to Table 15 in Example 1 hereinaftergiven). This is the reason why we have now finally decided thatcaprazene produced by us is a novel compound which is different from theCompound 10 in some part of the stereo-structure.

We have further succeeded in synthesizing 5″-amino-protected derivativesof caprazene by introducing, into the free amino group of caprazene ofthe formula (I) above, an alkoxycarbonyl group, for example,tert-butoxycarbonyl group (usually abbreviated as Boc), or anaralkyloxycarbonyl group, for example, benzyloxycarbonyl group, each ofwhich is conventionally used as an amino-protecting group in sugarchemistry.

According to a first aspect of this invention, therefore, there areprovided caprazene which is the compound represented by the followingformula (I)

wherein Me stands for methyl group, and a 5″-N-alkoxycarbonyl or5″-N-aralkyloxycarbonyl derivative thereof.

Further, according to a second aspect of this invention, there isprovided a process for the preparation of caprazene which is thecompound represented by the following formula (I)

wherein Me stands for methyl group, the process comprising hydrolyzingcaprazamycin A, B, C, D, E, F or G or a mixture of two or more ofcaprazamycins A to G in an aqueous solution of an acid at roomtemperature or under heating.

In the process according to the second aspect of this invention, it ispreferred that at least one of caprazamycins A to G is hydrolyzed in anaqueous acid solution, for example, in an aqueous acetic acid or anaqueous sulfuric acid or an aqueous hydrogen chloride solution.

The aqueous solution of an acid used for the acid hydrolysis ofcaprazamycins may be those of an organic acid, for example, acetic acidor n-propionic acid, or those of an inorganic acid, for example,hydrochloric acid or sulfuric acid. It is preferred to use an aqueousacetic acid solution containing acetic acid at a concentration of 50-90%(by weight) or a dilute aqueous hydrochloric acid solution containinghydrogen chloride (HCl) at a concentration of 3% by weight or less. Theacid hydrolysis reaction of caprazamycins may be carried out at roomtemperature, but it may also be effected at an elevated temperature of40-100° C.

After the finish of the hydrolysis reaction of caprazamycins, theresulting reaction solution is concentrated to give a syrupyconcentrate, to which is added acetone to deposit a precipitate, whichis then recovered by filtration. The resulting solid is washed withacetone and dried, and thus caprazene of the formula (I) may berecovered as a colorless solid. The solid caprazene may be dissolved ina mixture of water-acetone and then deposited as crystals. Thephysicochemical properties of caprazene are shown in Example 1hereinafter given.

We have further proceeded with our investigations. Thus, we have foundthat caprazene is suspended in a mixture of water-dioxane (2:1) and tothe resulting suspension is added triethylamine, so that there may beobtained a homogeneous caprazene solution. We have further found thatthere can be produced 5″-N-t-butoxycarbonyl caprazene or5″-N-benzyloxycarbonyl caprazene by reacting caprazene present in theresulting homogeneous solution with di-t-butyl dicarbonate having thefollowing formula (X)

or N-(benzyloxycarbonyloxy)succinimide to cause t-butoxy-carbonylationor benzyloxycarbonylation reaction at the 5-amino group of the5-amino-5-deoxy-D-ribose moiety of caprazene.

The 5″-N-t-butoxycarbonylcaprazene or 5″-benzyloxy-carbonylcaprazenethus formed is suspended in tetrahydrofuran (THF), and to the resultingsuspension are then added triethylamine andN,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride as activator ofcarboxyl group to give a homogeneous reaction mixture, to which mixtureis then added an amine compound of the following general formula (XI)

R¹—NH₂  (XI)

wherein R¹ is a straight chain or a substantially straight chain alkylgroup of 5-21 carbon atoms, a straight chain or a substantially straightchain alkenyl group of 5-21 carbon atoms or a cycloalkyl group of 5-12carbon atoms, or R¹ is a phenyl group having a straight chain alkylgroup of 1-14 carbon atoms or a straight chain alkoxy group of 1-9carbon atoms or a cycloalkyl group of 5-12 carbon atoms at thepara-position of the phenyl group, so as to bring about an amidationreaction on the 2″′-carboxyl group of the caprazene with the aminecompound of the formula (XI), and thus there can be produced a5″-N-protected-caprazene-1″′-amide derivative represented by thefollowing general formula (IIa)

wherein Me stands for methyl group, R¹ has the same meaning as R¹ in theformula (XI) above and A¹ stands for t-butoxycarbonyl group (abbreviatedas Boc) or benzyloxycarbonyl group (abbreviated as Z). In addition, ithas been found that the protection of 5″-amino group of caprazene of theformula (I)can also be made by using any alkoxycarbonyl group or aralkyloxycarbonylgroup conventionally used as the amino-protecting group in sugarchemistry instead of the above-said t-butoxycarbonyl group orbenzyloxycarbonyl group.

The elimination of 5″-N-Boc group or 5″-N-Z group from the amidederivative of the formula (IIa) can be made when the 5″-N-protectedcaprazene-1″′-amide derivative is subjected to conventional method forthe elimination of the amino-protecting group in sugar chemistry, forexample, to hydrolysis with trifluoroacetic acid in methanol for theelimination of Boc group, or to hydrogenolysis for the elimination of Zgroup, thereby affording a caprazene-1″″-amide derivative having thefollowing general formula (IIb)

wherein Me and R¹ have the same meanings as defined above. Thecaprazene-1″′-amide derivative of the general formula (IIb), when bereacted with trifluoroacetic acid, hydrochloric acid, sulfuric acid orphosphoric acid, will give the corresponding acid addition salt of theamide derivative of the formula (IIb) which is soluble in water.

We have found that a caprazene-1″′-amide derivative of the generalformula (IIb) above and 5″-N-Boc- or 5″-N-Z-protected derivative thereofhave antibacterial activities against various bacteria includingtubercle bacillus.

According to a third aspect of this invention, therefore, there areprovided a caprazene-1″′-amide derivative and its 5″-N-alkoxycarbonyl oraralkyloxycarbonyl derivative which are each presented by the followinggeneral formula (II)

wherein Me is methyl group, R¹ is a straight chain or a substantiallystraight chain alkyl group of 5-21 carbon atoms, a straight chain or asubstantially straight chain alkenyl group of 5-21 carbon atoms or acycloalkyl group of 5-12 carbon atoms, or R¹ is a phenyl group having astraight chain alkyl group of 1-14 carbon atoms or a straight chainalkoxy group of 1-9 carbon atoms or a cycloalkyl group of 5-12 carbonatoms at the para-position of the phenyl group, A is hydrogen atom or Ais an amino-protecting group including an alkoxycarbonyl group,particularly tert-butoxycarbonyl group, or an aralkyloxycarbonyl group,particularly benzyloxycarbonyl group, or a pharmaceutically acceptableacid addition salt thereof.

The caprazene-1″′-amide derivative of the general formula (II) includes(i) a caprazene-1″′-amide derivative where A is hydrogen atom and R¹ isan alkyl group, alkenyl group or cycloalkyl group as defined above, and(ii) a caprazene-1″′-amide derivative where A is hydrogen atom and R¹ isa phenyl group having an alkyl group, alkoxy group or cycloalkyl groupat the para-position as defined above.

In the caprazene-1″′-amide derivatives of the formula (II), the straightchain alkyl group of 5-21 carbon atoms as R¹ may be those exemplified inthe following Table 1.

TABLE 1 Alkyl group Formula Name C₅H₁₁— Pentyl(Amyl) C₆H₁₃— Hexyl C₇H₁₅—Heptyl C₈H₁₇— Octyl C₉H₁₉— Nonyl C₁₀H₂₁— Decyl C₁₁H₂₃— Undecyl C₁₂H₂₅—Dodecyl C₁₃H₂₇— Tridecyl C₁₄H₂₉— Tetradecyl C₁₅H₃₁— Pentadecyl C₁₆H₃₃—Hexadecyl C₁₇H₃₅— Heptadecyl C₁₈H₃₇— Octadecyl C₁₉H₃₉— Nonadecyl C₂₀H₄₁—Icocyl C₂₁H₄₃— Henicosyl

Substantially straight chain alkyl group of 5-21 carbon atoms as R¹defined above may be a (C₅-C₂₁) alkyl group having 1-3 methyl groups,1-3 ethyl groups or 1-3 n-propyl groups as substituted in the length ofthe alkyl chain or at the terminal carbon atom of the alkyl chain, whichmay, for example, include 9-methyl-undecyl group —(CH₂)₈CH(CH₃)CH₂CH₃ or10-methyl-undecyl group —(CH₂)₉CH(CH₃)₂.

Straight chain alkenyl group of 5-21 carbon atoms as R¹ defined abovemay be pentenyl group, hexenyl group, heptenyl group, octenyl group,nonenyl group, decenyl group, undecenyl group, dodecenyl group,tridecenyl group, tetradecenyl group, pentadecenyl group, hexadecenylgroup, heptadecenyl group, octadecenyl group, nonadecenyl group oricosenyl group. The double bond of the alkenyl group may be positionedin the length of the alkenyl chain or at the α-carbon atom or ω-carbonatom.

Cycloalkyl group of 5-12 carbon atoms as R¹ defined above may becyclopentyl group, cyclohexyl group, cycloheptyl group, cyclooctylgroup, cyclononyl group, cyclodecyl group, cycloundecyl group orcyclododecyl group. There may be substituted 1-3 methyl groups or 1-3ethyl groups as the substituents on the cycloalkane ring.

Concrete examples of a phenyl group having an alkyl group, alkoxy groupor cycloalkyl group at the para-position defined above for R¹ are shownin the column of R¹ in the Table 2-2 given hereinafter.

Concrete examples of a caprazene-1″′-amide derivative of the followingformula (IIb) which is included within the5″-N-unprotected-caprazene-1″′-amide derivative of the general formula(II) according to the third aspect of this invention are shown in thefollowing Table 2-1 and Table 2-2 together with their specific rotationdata.

TABLE 2-1 (IIb)

Specific rotation [α]_(D) Compound R¹ group in the formula (c 0.5, incode name (IIb) water) Compound II-A —(CH₂)₅CH₃ [α]_(D) ²⁰ +70° CompoundII-B —(CH₂)₆CH₃ [α]_(D) ²¹ +72° Compound II-C —(CH₂)₇CH₃ [α]_(D) ²⁰ +72°Compound II-D —(CH₂)₈CH₃ [α]_(D) ²⁰ +73° Compound II-E —(CH₂)₉CH₃[α]_(D) ²¹ +72° Compound II-F —(CH₂)₁₀CH₃ [α]_(D) ²⁰ +73° Compound II-G—(CH₂)₁₁CH₃ [α]_(D) ²¹ +72° Compound II-H —(CH₂)₁₂CH₃ [α]_(D) ²⁰ +72°Compound II-I —(CH₂)₁₃CH₃ [α]_(D) ²⁰ +68° Compound II-J —(CH₂)₁₄CH₃[α]_(D) ²¹ +66° Compound II-K —(CH₂)₁₅CH₃ [α]_(D) ²⁰ +67° Compound II-L—(CH₂)₁₆CH₃ [α]_(D) ²⁰ +67° Compound II-M —(CH₂)₁₇CH₃ [α]_(D) ²⁰ +66°Compound II-N —(CH₂)₁₈CH₃ [α]_(D) ²⁰ +60° Compound II-O —(CH₂)₁₉CH₃[α]_(D) ²⁰ +60° Compound II-P —(CH₂)₂₀CH₃ [α]_(D) ²⁰ +60° Compound II-QCyclododecyl group [α]_(D) ²⁰ +71° Compound II-R Oleyl group [α]_(D) ²⁰+64° —(CH₂)₈CH═CH(CH₂)₇CH₃ (cis form)

TABLE 2-2 (IIb)

Specific rotation [α]_(D) ²² Compound code R¹ group in the formula (c0.5, in name (IIb) methanol) compound II-1

+81° Compound II-2

+80° Compound II-3

+78° Compound II-4

+76° Compound II-5

+74° Compound II-6

+73° Compound II-7

+72° Compound II-8

+71° Compound II-9

+69° Compound II-10

+67° Compound II-11

+67° Compound II-12

+66° Compound II-13

+66° Compound II-14

+64° Compound II-15

+80° Compound II-16

+80° Compound II-17

+80° Compound II-18

+81° Compound II-19

+77° Compound II-20

+78° Compound II-21

+76° Compound II-22

+76° Compound II-23

+74° Compound II-24

+76°

Concrete examples of a caprazene-1″′-amide derivative of the followingformula (IIa) which is included within the5″-N-protected-caprazene-1″′-amide derivative of the general formula(II) according to the third aspect of this invention are shown in thefollowing Table 3 together with their specific rotation data.

TABLE 3 (IIa)

Specific R¹ group in Amino-protecting rotation [α]_(D) ¹⁹ Compound codethe formula group (A′) in (c 0.5, in name (IIa) formula (IIa)chloroform) Compound —(CH₂)₁₁CH₃ t-butoxycarbonyl +105° II-G-N-Boc groupCompound —(CH₂)₁₂CH₃ t-butoxycarbonyl +105° II-H-N-Boc group Compound—(CH₂)₁₃CH₃ t-butoxycarbonyl +103° II-I-N-Boc group Compound —(CH₂)₁₄CH₃t-butoxycarbonyl +103° II-J-N-Boc group

Test Example 1

Minimum growth inhibitory concentrations (mcg/ml) of caprazene-1″′-amidederivatives of the formula (II) against some of microorganisms weremeasured on an agar culture medium by a serial dilution method accordingto the standard method as provided by Japanese Society of Chemotherapy.For the cultivation of Mycobacterium smegmatis (one of anti-fastbacteria), however, there was used an agar culture medium with theaddition of 1% glycerine (the same applies to in the following tests).The results are shown in the following Table 4-1 and Table 4-2.

TABLE 4-1 Minimum growth inhibitory concentration (mcg/ml) Compound codename of against bacteria the test compound Staphylococcus MicrococcusMycobacterium (see Table 2, aureus luteus smegmatis Table 3) FDA209PFDA16 ATCC607 Compound II-A >100 3.13 100 Compound II-B 25 1.56 25Compound II-C 6.25 0.78 12.5 Compound II-D 6.25 0.39 3.13 Compound II-E25 0.39 0.78 Compound II-F 1.56 0.39 0.39 Compound II-G 3.13 0.39 0.39Compound II-H 3.13 0.78 <0.20 Compound II-I 3.13 0.78 0.78 Compound II-J1.56 0.78 1.56 Compound II-K 3.13 0.78 3.13 Compound II-L 1.56 1.56 3.13Compound II-M 6.25 0.78 3.13 Compound II-N 6.25 0.78 50 Compound II-O3.13 3.13 25 Compound II-P 6.25 3.13 50 Compound II-Q 100 0.20 3.13Compound II-R 1.56 1.56 1.56 Compound II-G-N-Boc 50 50 50 CompoundII-H-N-Boc 25 12.5 25 Compound II-I-N-Boc 12.5 12.5 25 CompoundII-J-N-Boc 12.5 12.5 25

TABLE 4-2 Minimum growth inhibitory concentration (mcg/ml) Compound Codeagainst bacteria name of the test Staphylococcus Mycobacterium Tcompound aureus Micrococcus smegmatis (see Table 2, Table 3) FDA209Pluteus FDA16 ATCC607 Compound II-1 >100 3.13 100 Compound II-2 100 3.1350 Compound II-3 25 3.13 12.5 Compound II-4 12.5 0.78 6.25 Compound II-53.13 0.2 3.13 Compound II-6 1.56 0.39 0.78 Compound II-7 1.56 <0.20<0.20 Compound II-8 3.13 0.39 0.78 Compound II-9 3.13 0.39 1.56 CompoundII-10 3.13 <0.20 3.13 Compound II-11 3.13 0.39 3.13 Compound II-12 3.130.78 6.25 Compound II-13 3.13 1.56 12.5 Compound II-14 6.25 3.13 25Compound II-15 100 3.13 50 Compound II-16 50 3.13 25 Compound II-17 251.56 12.5 Compound II-18 12.5 0.78 6.25 Compound II-19 6.25 0.78 3.13Compound II-20 6.25 0.39 1.56 Compound II-21 3.13 0.20 0.78 CompoundII-22 3.13 0.39 0.78 Compound II-23 1.56 0.39 0.78 Compound II-24 12.50.39 1.56

The process for the preparation of a caprazene-1″′-amide derivative ofthe formula (II) according to the third aspect of this invention is nowexplained.

At first, caprazene of the formula (I) is suspended in a mixture ofwater-dioxane, and to the resulting suspension is added triethylamine toprepare a homogeneous solution of caprazene. To the resulting caprazenesolution, is added an alkoxycarbonylating reagent or anaralkyloxycarbonylating reagent which is conventionally used accordingto the amino-protecting technique well known in the sugar chemistry, andthe reaction intended is conducted at room temperature. Thus, there isproduced in the resulting reaction solution a 5″-N-alkoxycarbonyl- or5″-N-aralkyloxycarbonyl-caprazene. The reaction solution is concentratedand the resulting solid residue is washed with ethyl acetate and thendried, and thus there is afforded the desired 5″-N-alkoxycarbonyl- or5″-N-aralkyloxycarbonyl-caprazene as a solid.

Then, 5″-N-alkoxycarbonyl- or 5″-N-aralkyloxycarbonyl-caprazene iseither dissolved in pyridine to give a solution or is suspended intetrahydrofuran (THF) to give a suspension and triethylamine is added tothe suspension. In the said pyridine solution or THF suspension of the5″-N-protected caprazene, an amine compound R¹—NH₂ of the formula (XI)above is reacted with the carboxyl group at the 2″′-position of the5″-N-protected-caprazene according to the usual method for the amidationof carboxylic acid. For the amidation reaction, it is convenient to addto the reaction system N,N-bis-(2-oxo-3-oxazolidinyl)-phosphinicchloride as an activator of the carboxyl group, and then to conduct thereaction at room temperature.

The resulting amidation reaction solution is concentrated, and theresulting syrupy concentrate is extracted with chloroform and then theresulting chloroform extract in the form of a solution is washed withwater and then concentrated, thus to give a residue containing thedesired 5″-N-protected-caprazene-1″′-amide derivative. The residue isdissolved in chloroform and the resulting solution is purified bysubjecting it to a silica-gel column chromatography which is developedwith a mixed solvent of chloroform-methanol (10:1). Eluate fractionscontaining the object product from the silica-gel column are collectedand the fractions collected are concentrated to afford a5″-N-protected-caprazene-1″′-amide derivative of the general formula(II) as a solid.

Further, the elimination of the 5″-N-protecting group from the resulting5″-N-protected-caprazene-1″′-amide derivative can be achieved bytreating the said N-protected derivative in a usual manner for theelimination of amino-protecting group, thereby to produce the5″-N-unprotected-caprazene-1″′-amide derivative of the general formula(II). In order to eliminate Boc group as the amino-protecting group, itis convenient, as described before, to dissolve the5″-N-protected-caprazene-1″′-amide derivative in methanol containing 80%trifluoroacetic acid (TFA) and then to stir the resulting solution atroom temperature. The resulting reaction solution from the eliminationof the amino-protecting group is concentrated to give a syrupyconcentrate, to which diethyl ether is added to deposit a precipitatewhich is then filtered, washed with diethyl ether and dried, thereby toyield the 5″-N-unprotected-caprazene-1″′-amide derivative of the generalformula (II) in the form of an addition salt of bis-trifluoroacetic acidas a solid.

We have further made another investigation. Thus, a5″-N-protected-caprazene as produced in the synthesis ofcaprazene-1″′-amide derivative of the general formula (II) above isdissolved in pyridine, and in the resultant pyridine solution, analcohol of the following general formula (XII)

R²—OH  (XII)

wherein R² is a straight chain or a substantially straight chain alkylgroup of 5-21 carbon atoms or a straight chain or a substantiallystraight chain alkenyl group of 5-21 carbon atoms or an alkynyl group of5-21 carbon atoms, is reacted with the 5″-N-protected caprazene at roomtemperature in the presence of N,N-bis(2-oxo-3-oxazolidinyl)phosphinicchloride as added (as activator of carboxyl group). Thus, there occursthe esterification reaction between the 2″′-carboxyl group of the5″-N-protected caprazene and the alcohol of the formula (XII), toproduce a 5″-N-protected caprazene-1″′-ester derivative represented bythe following general formula (IIIa)

wherein R² has the same meaning as defined above and A¹ is anamino-protecting group.

The resulting reaction solution containing the 5″-N-protectedcaprazene-1″′-ester derivative so produced is concentrated and theresulting syrupy concentrate is extracted with chloroform and then thesaid chloroform solution is washed with water and concentrated. Theresulting residue is dissolved in chloroform and the resultantchloroform solution is purified by subjecting it to a silica-gel columnchromatography with the development with a mixed solvent ofchloroform-methanol (10:1). The eluate fractions containing the desiredproduct from the column are concentrated to give a5″-N-protected-caprazene-1″′-ester derivative of the formula (IIIa) as asolid. The 5″-N-protected-caprazene-1″′-ester derivative of the formula(IIIa) has been found to have an antibacterial activity againstbacteria.

It has further been found that when the5″-N-protected-caprazene-1″′-ester derivative of the formula (IIIa) istreated in the same manner as described above, for the elimination ofthe amino-protecting group, the 5″-N-protecting group (A¹) can beeliminated, thus to produce a caprazene-1″′-ester derivative representedby the following general formula (IIIb)

wherein R² has the same meaning as defined above. Thecaprazene-1″′-ester derivative of the formula (IIIb), too, has beenfound to have an antibacterial activity against bacteria.

According to a fourth aspect of this invention, therefore, there areprovided a caprazene-1″′-ester derivative and a 5″-N-alkoxycarbonyl or a5″-N-aralkyloxycarbonyl derivative thereof which are each represented bythe following general formula (III):

wherein Me is methyl group, R² is a straight chain or a substantiallystraight chain alkyl group of 5-21 carbon atoms or a straight chain or asubstantially straight chain alkenyl group of 5-21 carbon atoms or analkynyl group of 5-21 carbon atoms and A is hydrogen atom or anamino-protecting group which is an alkoxycarbonyl group, particularlytert-butoxycarbonyl group, or an aralkyloxycarbonyl group, particularlybenzyloxycarbonyl group, or a pharmaceutically acceptable acid additionsalt thereof.

In the 5″-N-unprotected or 5″-N-protected caprazene-1″′-ester derivativehaving the general formula (III), a straight chain or a substantiallystraight chain alkyl or alkenyl group of 5-21 carbon atoms for R² eachmay be the same as the alkyl or alkenyl group defined for R² in thecaprazene-1″′-amide derivative having the general formula (II),respectively. An alkynyl group of 5-21 carbon atoms for R² may bepentynyl group, hexynyl group, heptynyl group, octynyl group, nonynylgroup, decynyl group and so on.

Concrete examples of the caprazene-1″′-ester derivative of the followingformula (IIIb) which are included within the 5″-N-unprotectedcaprazene-1″′-ester derivative of the general formula (III) according tothe fourth aspect of this invention are shown in the following Table 5together with their compound codes and specific rotation data.

TABLE 5 (IIIb)

Specific Compound code R² group in the formula rotation [α]_(D) ¹⁹ name(IIIb) (c 0.5, in water) Compound III-AA —(CH₂)₉CH₃ +46° Compound III-BB—(CH₂)₁₂CH₃ +50° Compound III-CC —(CH₂)₁₇CH₃ +44° Compound III-DD—(CH₂)₁₀—CH═CH—CH₂—CH₃ +42° Compound III-EE —CH₂—CH═CH—(CH₂)₈—CH₃ +48°Compound III-FF —(CH₂)₉—CH═CH₂ +48° Compound III-GG—(CH₂)₂—C≡C—(CH₂)₅—CH₃ +40°

Test Example 2

Minimum growth inhibitory concentrations (mcg/ml) of some of thecaprazene-1″′-ester derivative of the formula (III) against some ofmicroorganisms were measured on an agar culture medium by a serialdilution method according to the standard method as provided by JapaneseSociety of Chemotherapy. The results obtained are shown in the followingTable 6.

TABLE 6 Minimum growth inhibitory concentration (mcg/ml) Compound codeagainst bacteria name of the test Staphylococcus Mycobacterium compound(see aureus Micrococcus smegmatis Table 5) FDA209P luteus FDA16 ATCC607Compound III-AA 12.5 1.56 12.5 Compound III-BB 3.13 3.13 12.5 CompoundIII-CC 12.5 6.25 >100 Compound III-DD 6.25 1.56 25 Compound III-EE 6.251.56 12.5 Compound III-FF 25 1.56 12.5 Compound III-GG 50 6.25 25

The process for the preparation of a caprazene-1″′-ester derivative ofthe formula (III) according to the fourth aspect of this invention isnow explained.

First of all, as explained in the third aspect of this invention, a5″-N-alkoxycarbonyl- or 5″-N-aralkyloxycarbonyl-caprazene is prepared.Then, the 5″-N-alkoxycarbonyl- or 5″-N-aralkyloxycarbonyl-caprazene isdissolved in pyridine, and in the resulting pyridine solution, analcohol compound of the formula (XII) above is reacted with the2″′-carboxyl group of the 5″-N-protected caprazene according to theusual method for the esterification of carboxylic acids. Theesterification reaction is conveniently carried out in the presence ofN,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride as added at roomtemperature.

The resulting esterification reaction solution is concentrated, and theresulting concentrate is extracted with chloroform and the resultingchloroform extract is washed with water and then concentrated, to give aresidue containing the desired 5″-N-protected caprazene-1″′-esterderivative. The residue is dissolved in chloroform and the resultingchloroform solution is purified by subjecting it to a silica-gel columnchromatography with the development with a mixed solvent ofchloroform-methanol. The eluate fractions containing the desired productfrom the chromatography are collected and concentrated, thus to yieldthe desired 5″-N-protected caprazene-1″′-ester derivative of the formula(IIIa) as a solid.

The elimination of the 5″-N-amino-protecting group can be achieved bytreating the 5″-N-protected caprazene-1″′-ester derivative by the usualmethod for the elimination of the amino-protecting group, to produce a5″-N-unprotected caprazene-1″′-ester derivative of the general formula(IIIb). In case where 5″-N-protecting group is Boc group, it isconvenient to eliminate Boc group by dissolving the 5″-N-protectedcaprazene-1″′-ester derivative in methanol containing 80% TFA andstirring the resultant solution at room temperature. The resultingreaction solution is concentrated, and to the concentrate is addeddiethyl ether to deposit a precipitate, and the precipitate is filtered.The solid precipitate separated is washed with diethyl ether and thendried, thus to afford a 5″-N-unprotected caprazene-1″′-ester derivativeof the general formula (IIIb) as a solid.

We have made further experiments on an alkaline hydrolysis of acaprazamycin at room temperature by adding an aqueous solution of aninorganic base, for example, aqueous ammonia solution or a diluteaqueous sodium hydroxide solution to an N,N-dimethylformamide solutionof caprazamycin A, B or C. As a result, it has been found that thealkaline hydrolysis of caprazamycin A, B or C gives the compoundrepresented by the following formula (IV)

wherein Me is methyl group, and the compound is successfully isolated asa colorless solid. The crystallization of this solid from a mixture ofwater-methanol could give colorless crystals of the said compound[melting point 205-206° C. (with decomposition)].

We have now decided that the compound of the formula (IV) thus isolatedhas the steric chemical structure of the formula (IV) given above bymeasuring the physicochemical properties and NMR data of said compoundof formula (IV) and by analyzing further this compound by X-ray powderdiffractometry.

Further, by taking the physicochemical properties, ¹H-NMR data and¹³C-NMR data of said compound of formula (IV) together intoconsideration, we have judged it to be a novel compound and designatedit as caprazol.

In addition, comparison has been made between caprazol of the formula(IV) of this invention and the aforesaid Compound II and Compound 12both of which have sulfuric acid group —SO₃H and which are given bytheir planer structural formulae in the above literature “The Journal ofOrganic Chemistry”, Vol. 57, No. 24, pp. 6397-6399 and 6402 and theirsteric structures are unknown yet. That is, when comparing the ¹³C-NMRdata (Table III) and ¹H-NMR data (Table IV) of Compounds II and 12 withthe ¹³C-NMR data and ¹H-NMR data (refer to Table 18 of Example 5 givenlater) of caprazol of this invention, it appears that the numerical dataof the former are not necessarily consistent in part with those of thelatter. Judging from this comparison, we have concluded that caprazol asprepared by us is different in some part of its steric structure and inthe presence or absence of sulfuric acid group, from Compounds 11 and12, and thus that caprazole is a novel compound.

According to a fifth aspect of this invention, therefore, there areprovided caprazol which is the compound represented by the followingformula (IV)

wherein Me is methyl group, and a 5″-N-alkoxycarbonyl and a5″-N-aralkyloxycarbonyl derivative thereof.

Further, according to a sixth aspect of this invention, there isprovided a process for the preparation of caprazol represented by thefollowing formula (IV)

which comprises subjecting caprazamycin A, B, C, D, E, F or G or amixture of at least two of caprazamycins A to G to a hydrolysis in anaqueous solution of an inorganic base at room temperature or underheating.

In the process according to the sixth aspect of this invention, it ispreferred that at least one of caprazamycins A to G is hydrolyzed in anaqueous ammonia solution containing 15-30% by weight of ammonia (NH₃) ata temperature of about 40° C. or lower.

The alkaline hydrolysis reaction of caprazamycins using an aqueousdilute sodium hydroxide or potassium hydroxide solution may be carriedout at room temperature or may also be carried out at an elevatedtemperature of 40-80° C.

After the finish of the alkaline hydrolysis reaction of caprazamycins,the resulting reaction solution is post-treated by filtering off theinsolubles therefrom, concentrating the resulting filtrate, washing theresulting solid residue with acetone and drying the resulting residue,and thus there can be recovered caprazol of the formula (IV) as acolorless solid. The solid caprazol thus recovered may be dissolved in awater-methanol mixture and then crystallized to afford the desiredsubstance as crystals. The physicochemical properties of caprazol areshown in Example 5 given hereinafter.

We have further proceeded our investigations. Thus, we have found that5″-N-t-butoxycarbonylcaprazol or 5″-N-benzyloxy-carbonylcaprazol may beproduced by a process comprising the steps of dissolving caprazol in asolution of dioxane in water and reacting caprazol in the resultantaqueous solution with triethylamine and di-t-butyl dicarbonate orN-(benzyloxycarbonyloxy)succinimide, so that the 5-amino group of the5-amino-5-deoxy-D-ribose moiety of caprazol is t-butoxycarbonylated orbenzyloxycarbonylated.

We have further succeeded in synthesizing a 5″-N-protected derivative ofcaprazol, generically, by introducing into the free amino group at the5″-position of caprazol of the formula (IV) an alkoxycarbonyl group, forexample, tert-butoxycarbonyl group (usually abbreviated as Boc), or anaralkyloxycarbonyl group, for example, benzyloxycarbonyl group, which isconventionally used as an amino-protecting group in the sugar chemistry.

There may be produced a 5″-N-protected caprazol-1″′-amide derivativerepresented by the following general formula (Va)

wherein Me stands for methyl group, R³ is a straight chain or asubstantially straight chain alkyl group of 5-21 carbon atoms, astraight chain or a substantially straight chain alkenyl group of 5-21carbon atoms or a cycloalkyl group of 5-12 carbon atoms, and A¹ standsfor t-butoxycarbonyl group (sometimes abbreviated as Boc) orbenzyloxycarbonyl group (sometimes abbreviated as Z), by conducting aprocess comprising the steps of dissolving 5″-N-t-butoxycarbonyl- orbenzyloxycarbonyl-caprazol in N,N-dimethylformamide, adding to theresulting solution triethylamine andN,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride, successively, andsubjecting the resulting solution to a reaction with an amine compoundof the following general formula (XIII)

R³—NH₂  (XIII)

wherein R³ has the same meaning as defined in the formula (Va) above, inthe presence of the said phosphinic chloride as added, so that there canoccur the desired amidation reaction between the 2″′-carboxyl group ofcaprazol and the amine compound of the formula (XIII). We have furtherfound that the protection of 5″-amino group of caprazol of the formula(IV) may also be made if the t-butoxycarbonyl group or benzyloxycarbonylgroup used above is replaced by any other alkoxycarbonyl group oraralkyloxycarbonyl group which is conventionally employed asamino-protecting group in the sugar chemistry.

In cases where the 5″-N-protected caprazol-1″′-amide derivative of theformula (Va) above contains Boc group as amino-protecting group, the5″-N-Boc group may be eliminated from the amide derivative of theformula (Va) by subjecting the said compound to a method for theelimination of the amino-protecting group conventionally employed in thesugar chemistry, for example, to a hydrolysis with trifluoroacetic acidin methanol, whereby there can be produced a caprazol-1″′-amidederivative represented by the following general formula (Vb)

wherein Me and R³ have the same meanings as defined above. Thecaprazol-1″′-amide derivative of the general formula (Vb) thus obtained,if it be reacted with trifluoroacetic acid, hydrochloric acid, sulfuricacid or phosphoric acid, can afford the corresponding acid addition saltof the amide derivative of the formula (Vb), which is soluble in water.

Further, we have now found that the caprazol-1″′-amide derivative of thegeneral formula (Vb) above and its 5″-N-Boc- or 5″-N-Z-protectedderivative, namely 5″-N-protected caprazol-1″′-amide derivative of thegeneral formula (Va), have antibacterial activities against a variety ofbacteria, including tubercle bacillus.

According to a seventh aspect of this invention, therefore, there areprovided a caprazol-1″′-amide derivative and its 5″-N-alkoxycarbonyl- oraralkyloxycarbonyl derivatives which are represented by the followinggeneral formula (V)

wherein Me is methyl group, R³ is a straight chain or a substantiallystraight chain alkyl group of 5-21 carbon atoms, a straight chain or asubstantially straight chain alkenyl group of 5-21 carbon atoms or acycloalkyl group of 5-12 carbon atoms, and A is hydrogen atom or analkoxycarbonyl group, particularly tert-butoxycarbonyl group, or anaralkyloxycarbonyl group, particularly benzyloxycarbonyl group as theamino-protecting group, or a pharmaceutically acceptable acid additionsalt thereof.

In the 5″-N-unprotected or -protected caprazol-1″′-amide derivative ofthe general formula (V) according to the seventh aspect of thisinvention, an alkyl group, alkenyl group and cycloalkyl group for R³ maybe the same as the alkyl group, alkenyl group and cycloalkyl group forR¹ present in the 5″-N-unprotected or -protected caprazene-1″′-amidederivative of the general formula (II) according to the third aspect ofthis invention, respectively.

Concrete examples of a caprazol-1″′-amide derivative of the followingformula (Vb) which are included within the 5″-N-unprotected or-protected caprazol-1″′-amide derivative of the general formula (V)according to the seventh aspect of this invention are shown in thefollowing Table 7 together with their Compound code names and specificrotation data.

TABLE 7 (Vb)

Specific rotation [α]_(D) ¹⁹ Compound code R³ group in the (c 0.5, inname formula (Vb) methanol) Compound V-A —(CH₂)₅CH₃ +15° Compound V-B—(CH₂)₆CH₃ Compound V-C —(CH₂)₇CH₃ +15° Compound V-D —(CH₂)₈CH₃ CompoundV-E —(CH₂)₉CH₃ +12° Compound V-F —(CH₂)₁₀CH₃ +12° Compound V-G—(CH₂)₁₁CH₃ +12° Compound V-Q Cyclododecyl group +35° Compound V-R Oleylgroup +14° —(CH₂)₈CH═CH(CH₂)₇CH₃ (cis-form)

Test Example 3

Minimum growth inhibitory concentrations (mcg/ml) of some of thecaprazol-1″′-amide derivative of the formula (V) against a variety ofmicroorganisms were measured on an agar medium by a serial dilutionmethod according to the standard method as provided by Japanese Societyof Chemotherapy. The results obtained are shown in the following Table8.

TABLE 8 Minimum growth inhibitory concentration (mcg/ml) Compound codeagainst bacteria name of the test Staphylococcus Mycobacterium compoundaureus Micrococcus smegmatis (see Table 7) FDA209P luteus FDA16 ATCC607Compound V-A 50 50 12.5 Compound V-B Compound V-C 25 25 6.25 CompoundV-D Compound V-E 25 25 6.25 Compound V-F 12.5 25 6.25 Compound V-G 12.525 6.25 Compound V-Q Compound V-R 12.5 25 6.25

The process for the preparation of a caprazol-1″′-amide derivative ofthe formula (V) is now explained.

Thus, caprazol of the formula (IV) is dissolved in water, and to theresulting aqueous solution of caprazol is added an alkoxycarbonylatingreagent or an aralkyloxycarbonylating reagent conventionally usedaccording to the amino-protecting technique well-known inorganicchemistry, desirably in the form of its solution in an organic solventsuch as dioxane, together with triethylamine. The reaction intended isthen effected at room temperature. There is produced a5″-N-alkoxycarbonyl- or 5″-N-aralkyloxycarbonyl-caprazol in theresulting reaction solution. An aqueous ammonia solution is added to thereaction solution and the resulting solution is concentrated under areduced pressure. The resulting solid residue is dried under a reducedpressure, thus affording the desired 5″-N-alkoxycarbonyl- or5″-N-aralkyloxycarbonyl-caprazol in the form of solid.

Subsequently, the 5″-N-alkoxycarbonyl- or 5″-N-aralkyloxycarbonyl-caprazol is dissolved in N,N-dimethylformamide, and to theresultant solution is added triethylamine, thus to give a homogeneoussolution of the 5″-N-protected caprazol. An amine compound R³—NH₂ of theformula (XIII) given above is reacted with the 2″′-carboxyl group of the5″-N-protected caprazol in the resulting solution in accordance with anyconventional method for amidation of the carboxylic acid. For theamidation reaction intended, it is convenient to carry out the reactionin the presence of N,N-bis(2-oxo-3-oxazolidinyl)phosphinic chlorideadded, at room temperature.

The resulting amidation reaction solution is concentrated, and thesyrupy concentrate obtained is extracted with chloroform and then theresulting chloroform extract is washed with water, dried andconcentrated to dryness, thus affording a solid residue which contains a5″-N-protected caprazol-1″′-amide derivative desired. The residue isdissolved in chloroform and the resultant solution is purified bysubjecting it to a silica-gel column chromatography with developmentwith a mixed solvent of chloroform-methanol-concentrated aqueous ammonia(4:1:0.1). Active fractions of the eluate from the silica-gel column arecollected and concentrated, whereby there can be yielded the desired5″-N-protected caprazol-1″′-amide derivative of the general formula (V)as a solid.

Further, the 5″-amino-protecting group can be eliminated by treating theresulting 5″-N-protected caprazol-1″′-amide derivative in accordancewith the conventional technique for the elimination of amino-protectinggroup, thereby producing the 5″-N-unprotected caprazol-1″′-amidederivative of the general formula (V). As explained above, for thepurpose of the elimination of the 5″-amino-protecting group, Boc, it isconvenient to dissolve the 5″-N-protected caprazol-1″′-amide derivativein methanol containing 80% trifluoroacetic acid (TFA) and to stir theresulting solution at room temperature. The resulting reaction solutionfrom the elimination of the amino-protecting group is concentrated, andto the resulting syrupy concentrate is added diethyl ether to deposit aprecipitate which is recovered by filtration. The precipitate thusrecovered is washed with diethyl ether and then dried, and thus therecan be afforded the desired 5″-N-unprotected caprazol-1″′-amidederivative of the general formula (V) in the form of an addition salt ofbis-trifluoroacetic acid as a solid.

We have further made a different study. It started with using the5″-N-t-butoxycarbonylcaprazol which was prepared in the synthesis of thecaprazol-1″′-amide derivatives of the general formula (V) according tothe seventh aspect of this invention. Thus, the5″-N-t-butoxycarbonylcaprazol was dissolved in N,N-dimethylformamide,and to the resultant solution were added, in order,anhydrous(±)10-camphorsulfonic acid (as acid catalyst) anddimethoxymethane, and the reaction was effected at room temperature.Thus, it has been found that by the reaction, the 2′- and 3′-hydroxylgroups and the 2″- and 3″-hydroxyl groups each of the5″-N-t-butoxycarbonylcaprazol are protected with isopropylidene group(═C(CH₃)₂; a known hydroxyl-protecting group), to produce5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazolrepresented by the following formula (XIV)

(see Example 9 (a) given hereinafter). It has also been found that whenthe caprazol-N,O-protected derivative of the formula (XIV) is dissolvedin N,N-dimethylformamide, and to the resultant solution are added, inorder, triethylamine and an amine compound of the formula (XIII) above,and when the subsequent amidation reaction is conducted at roomtemperature in the same manner as that in the preparation of thecaprazol-1″′-amide derivative of the general formula (V) according tothe seventh aspect of this invention, there can be produced5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-amidederivative which is represented by the following general formula (XV)

wherein R³ is a straight chain or a substantially straight chain alkylgroup of 5-21 carbon atoms or a straight chain or a substantiallystraight chain alkenyl group of 5-21 carbon atoms or a cycloalkyl groupof 5-12 carbon atoms (see Example 9 (b) hereinafter given). Theresulting amidation reaction solution containing the N,O-protectedcaprazol-1″′-amide derivative of the formula (XV) is concentrated todryness, and the residue obtained is extracted with chloroform and thenthe chloroform extract is washed with water, dried and concentrated todryness. The resulting solid residue is dissolved in chloroform and theresulting solution is purified by subjecting it to a silica-gel columnchromatography by development with a mixed solvent ofchloroform-methanol (50:1). The eluate fractions containing the desiredproduct from the silica-gel column are collected and concentrated, andthus there can be recovered the N,O-protected caprazol-1″′-amidederivative of the formula (XV).

Subsequently, the N,O-protected caprazol-1″′-amide derivative of theformula (XV) is dissolved in dichloromethane, and to the resultantsolution are added 4-dimethylaminopyridine and an acid chloride of thefollowing formula (XVI)

Cl—CO—R⁴  (XVI)

wherein R⁴ is a straight chain or a substantially straight chain alkylgroup of 5-21 carbon atoms or a straight chain or a substantiallystraight chain alkenyl group of 5-21 carbon atoms or an alkynyl group of5-21 carbon atoms, and the reaction intended is conducted underice-cooling. Thus, the 3″′-hydroxyl group of the N,O-protectedcaprazol-1″′-amide derivative of the formula (XV) is acylated with theacid chloride of the formula (XVI), whereby there can be produced a5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-amide-3″′-esterderivative represented by the following general formula (XVII)

wherein R³ and R⁴ have the same meanings as defined above.

To the resulting acylation reaction solution containing theN,O-protected caprazol-1″′-amide-3″′-ester derivative of the formula(XVII) is added a small amount of methanol to decompose the residualreagent. Then the reaction solution is diluted with chloroform and theresulting solution is washed with an aqueous potassium hydrogen sulfateand water, and then the so washed solution is dried and concentrated todryness. The resulting residue is dissolved in chloroform and theresultant solution is purified by subjecting it to a silica-gel columnchromatography [at first, washing with chloroform, followed bydeveloping with chloroform-methanol (150:1)]. There can thus berecovered the derivative of the formula (XVII) by concentrating theeluate fractions containing the derivative of the formula (XVII).

Thereafter, the elimination of the 5″-t-butoxycarbonyl group and the twoisopropylidene groups (═C(CH₃)₂) can be effected by treating thederivative of the formula (XVII) with trifluoroacetic acid in methanol,thus to afford the caprazol-1″′-amide-3″′-ester derivative of thegeneral formula (VI) given below. The resulting reaction solution fromthe treatment with trifluoroacetic acid for the deprotection of theprotecting groups is then concentrated to dryness and the residue iswashed with diethyl ether, whereby an addition salt of trifluoroacetaticacid of the caprazol-1″′-amide-3″′-ester derivative of the formula (VI)can be recovered. The caprazol-1″′-amide-3″′-ester derivative of theformula (VI) has been found also to possess antibacterial activitiesagainst bacteria.

According to an eighth aspect of this invention, therefore, there isprovided a caprazol-1″′-amide-3″′-ester derivative represented by thefollowing general formula (VI)

wherein Me is methyl group, R³ is a straight chain or a substantiallystraight chain alkyl group of 5-21 carbon atoms or a straight chain or asubstantially straight chain alkenyl group of 5-21 carbon atoms or acycloalkyl group of 5-12 carbon atoms, R⁴ is a straight chain or asubstantially straight chain alkyl group of 5-21 carbon atoms or astraight chain or a substantially straight chain alkenyl group of 5-21carbon atoms or an alkynyl group of 5-21 carbon atoms, or apharmaceutically acceptable acid addition salt thereof.

Some typical examples of a caprazol-1″′-amide-3″′-ester derivative ofthe general formula (VI) according to the eighth aspect of thisinvention are shown in the following Table 9 together with the Compoundcode names and their specific rotation data.

TABLE 9 (VI)

Specific rotation [α]_(D) ²¹ Compound R⁴ group in (c 0.5, in code R³group in the formula the formula meth- name (VI) (VI) anol) Compound—(CH₂)₅CH₃ —(CH₂)₅CH₃ +6° VI-A Compound —(CH₂)₆CH₃ —(CH₂)₆CH₃ VI-BCompound —(CH₂)₇CH₃ —(CH₂)₇CH₃ +6° VI-C Compound —(CH₂)₈CH₃ —(CH₂)₈CH₃VI-D Compound —(CH₂)₉CH₃ —(CH₂)₉CH₃ +5° VI-E Compound —(CH₂)₁₀CH₃—(CH₂)₁₀CH₃ +6° VI-F Compound —(CH₂)₁₁CH₃ —(CH₂)₁₀—CH₃ +6° VI-G CompoundCyclododecyl group —(CH₂)₁₀—CH₃ +24°  VI-Q Compound—(CH₂)₈CH═CH(CH₂)₇CH₃ —(CH₂)₁₀—CH₃ +5° VI-R (cis-form)

Test Example 4

Minimum growth inhibitory concentrations (mcg/ml) of some of thecaprazol-1″′-amide-3″′-ester derivative of the formula (VI) against avariety of microorganisms were measured on an agar medium by a serialdilution method according to the standard method as provided by JapaneseSociety of Chemotherapy. The results obtained are shown in the followingTable 10.

TABLE 10 Minimum growth inhibitory concentration (mcg/ml) Compound codeagainst bacteria name of the test Staphylococcus Mycobacterium compound(see aureus Micrococcus smegmatis Table 9) FDA209P luteus FDA16 ATCC607Compound VI-A 25 25 12.5 Compound VI-B Compound VI-C 12.5 6.25 6.25Compound VI-D Compound VI-E 12.5 6.25 6.25 Compound VI-F 12.5 3.13 6.25Compound VI-G 25 3.13 6.25 Compound VI-Q 12.5 3.13 6.25 Compound VI-R 253.13 6.25

We have further made a different investigation. Thus, the5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazol of theformula (XIV) prepared as above is dissolved in dichloromethane, and tothe resultant solution are added 4-dimethylamino-pyridine and an acidchloride of the following formula (XVI)

Cl—CO—R⁴  (XVI)

wherein R⁴ is a straight chain or a substantially straight chain alkylgroup of 5-21 carbon atoms or a straight chain or a substantiallystraight chain alkenyl group of 5-21 carbon atoms or an alkynyl group of5-21 carbon atoms, and the reaction intended is effected underice-cooling. So, The 3″′-hydroxyl group of the N,O-protected caprazol ofthe formula (XIV) is acylated with the acid chloride of the formula(XVI), and thus there can be yielded a5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazol-3″′-esterderivative represented by the following general formula (XVIII)

wherein R⁴ has the same meaning as defined above.

To the resulting acylation reaction solution containing theN,O-protected caprazol-3″′-ester derivative of the formula (XVIII) isadded a small amount of methanol to decompose the residual reagent.Then, the resulting solution is diluted with chloroform and theresulting solution is washed with an aqueous potassium hydrogen sulfatesolution and water, in order, and the so washed solution is dried andconcentrated to dryness. Thus, there can be recovered theN,O-protected-3″′-ester derivative of the formula (XVIII) as a solid.

Subsequently, the N,O-protected-3″′-ester derivative of the formula(XVIII) is treated with trifluoroacetic acid in methanol, thereby toeliminate the 5″-t-butoxycarbonyl group (Boc) and the two isopropylidenegroups, and thus there can be produced a caprazol-3″′-ester derivativerepresented by the following general formula (XIX)

wherein R⁴ has the same meaning as defined above. The resulting reactionsolution containing the caprazol-3″′-ester derivative of the formula(XIX) from the deprotecting treatment with trifluoroacetic acid isconcentrated to dryness and the resulting residue is washed with diethylether, and thus there can be recovered an addition salt oftrifluoroacetic acid of a caprazol-3″′-ester derivative of the formula(XIX). The caprazol-3″′-ester derivative of the formula (XIX) has beenfound also to have antibacterial activities against bacteria.

In an another study, the N,O-protected caprazol of the formula (XIV)above is dissolved in N,N-dimethylformamide, and to the resultantsolution are added successively triethylamine andN,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride, and further added asan esterifying reagent an alkanol of the following formula (XX)

R⁷—OH  (XX)

wherein R⁷ is an alkyl group of 1-21 carbon atoms, and the reactionintended is conducted at room temperature. Thus, the 2″′-carboxyl groupof the N,O-protected caprazol of the formula (XIV) can be esterified toproduce a5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-esterderivative represented by the following general formula (XXI)

The resulting esterifying reaction solution is concentrated to drynessand the resulting residue is extracted with chloroform. The chloroformextract is washed with water, dried and concentrated to dryness, and theresulting residue is dissolved in chloroform. The chloroform solution ispurified by subjecting it to a silica-gel column chromatography withdevelopment with chloroform-methanol (50:1). The desired eluatefractions are collected and concentrated to dryness, and thus there canbe recovered the desired N,O-protected caprazol-1″′-ester derivative ofthe formula (XXI).

The N,O-protected caprazol-1″′-ester derivative of the formula (XXI) isthen dissolved in dichloromethane, and to the resulting solution areadded 4-dimethylaminopyridine and an acid chloride of the followingformula (XVI)

Cl—CO—R⁴  (XVI)

wherein R⁴ is a straight chain or a substantially straight chain alkylgroup of 5-21 carbon atoms or a straight chain or a substantiallystraight chain alkenyl group of 5-21 carbon atoms or an alkynyl group of5-21 carbon atoms. The reaction intended is effected wherein R⁴ is astraight chain or a substantially straight chain alkyl group of 5-21carbon atoms or a straight chain or a substantially straight chainalkenyl group of 5-21 carbon atoms or an alkynyl group of 5-21 carbonatoms. The reaction intended is effected under ice-cooling. Thus, the3″′-hydroxyl group of the N,O-protected caprazol-1″′-ester derivative ofthe formula (XXI) can be acylated with the acid chloride to produce a5″-N-t-butoxycarbonyl-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-ester-3″′-esterderivative represented by the following general formula (XXII)

wherein R⁴ and R⁷ have the same meanings as defined above.

To the resulting acylation reaction solution containing theN,O-protected caprazol-1″′-ester-3″′-ester derivative of formula (XXII)is added a small amount of methanol to decompose the residual reagent.Then, the solution is diluted with chloroform and the resulting dilutedsolution is washed with an aqueous potassium hydrogen sulfate solutionand water, and the solution thus washed is dried and concentrated todryness. The resulting residue is dissolved in chloroform and theresulting solution is purified by subjecting it to a silica-gel columnchromatography in the same manner as above. There can be recovered a1″′-ester-3″′-ester derivative of the formula (XXII) by concentratingthe eluate fractions containing the 1″′-ester-3″′-ester derivative ofthe formula (XXII).

Subsequently, for the purpose of deprotection of the 1″′-ester-3″′-esterderivative of the formula (XXII), the treatment of this derivative withtrifluoroacetic acid is carried out in methanol in the same manner asthat above-mentioned. Thus, the 5″-t-butoxycarbonyl group and the twoisopropylidene groups can be eliminated to produce acaprazol-1″′-ester-3″′-ester derivative of the following general formula(XXIII)

wherein R⁴ and R⁷ have the same meanings as defined above. The resultingreaction solution from the deprotection treatment with trifluoroaceticacid is concentrated to dryness, and the resulting residue is washedwith diethyl ether, and thus there can be recovered an addition salt oftrifluoroacetatic acid of the caprazol-1″′-ester-3″′-ester derivative ofthe formula (XXIII). The caprazol-1″′-ester-3″′-ester derivative of theformula (XXIII) as well as the caprazol-3″′-ester derivative of theformula (XIX) has been found also to have antibacterial activitiesagainst bacteria.

According to a ninth aspect of this invention, therefore, there isprovided a caprazol-3″′-ester derivative or acaprazol-1″′-ester-3″′-alkyl ester derivative which is represented bythe following general formula (VII)

wherein Me is methyl group, R⁴ is a straight chain or a substantiallystraight chain alkyl group of 5-21 carbon atoms or a straight chain or asubstantially straight chain alkenyl group of 5-21 carbon atoms or analkynyl group of 5-21 carbon atoms and R⁵ is hydrogen atom or an alkylgroup of 1-21 carbon atoms, or a pharmaceutically acceptable acidaddition salt thereof.

Some concrete examples of a caprazol-3″′-ester derivative or acaprazol-1″′-ester-3″′-ester derivative of the general formula (VII)according to the ninth aspect of this invention are shown in thefollowing Table 11 together with their compound code names and specificrotation data.

TABLE 11 (VII)

R⁵ group Compound in the Specific code R⁴ group in the formula rotationname formula (VII) (VII) [α]_(D) ²⁰ Compound —(CH₂)₅CH₃ —H +16°(c 0.5,VII-A in DMSO) Compound —(CH₂)₆CH₃ —H +16°(c 0.5, VII-B in DMSO)Compound —(CH₂)₇CH₃ —H +16°(c 0.5, VII-C in DMSO) Compound —(CH₂)₈CH₃ —H+17°(c 0.5, VII-D in DMSO) Compound —(CH₂)₉CH₃ —H +17°(c 0.5, VII-E inDMSO) Compound —(CH₂)₁₀CH₃ —H +17°(c 0.5, VII-F in DMSO) Compound—(CH₂)₁₀CH₃ —CH₃ +6°(c 1, in VII-G methanol) Compound Cyclododecyl group—H VII-Q Compound —(CH₂)₇CH═CH(CH₂)₇CH₃ —H +14°(c 0.5, VII-R (cis-form)in DMSO)

Test Example 5

Minimum growth inhibitory concentrations (mcg/ml) of some of thecaprazol-3″′-ester derivative or caprazol-1″′-ester-3″′-ester derivativeof the formula (VII) against a variety of microorganisms were measuredon an agar medium by a serial dilution method according to the standardmethod as provided by Japanese Society of Chemotherapy. The resultsobtained are shown in the following Table 12.

TABLE 12 Minimum growth inhibitory concentration (mcg/ml) Compound codeagainst bacteria name of the test Staphylococcus Mycobacterium compound(see aureus Micrococcus smegmatis Table 11) FDA209P luteus FDA16 ATCC607Compound VII-A 25 >50 1.56 Compound VII-B 12.5 >50 1.56 Compound VII-C12.5 >50 0.78 Compound VII-D 3.13 3.13 0.78 Compound VII-E 1.56 1.560.39 Compound VII-F 0.78 3.13 0.78 Compound VII-G >100 >100 50 CompoundVII-Q Compound VII-R 1.56 3.13 6.25

We have further proceeded with a different investigation. Thus, caprazolis treated with methylamine in an aqueous solution of caprazol of theformula (IV) at room temperature for a long period of time. It has beenfound that by this treatment reaction, the diazepinone ring moiety ofcaprazol can be opened to produce an uridine derivative of the followingformula (IX)

The resulting reaction solution from the reaction of caprazol withmethylamine is concentrated under a reduced pressure and dried, and theresulting residue is washed with a mixed solvent of chloroform-diethylether and dried. The solid thus obtained is dissolved in water. Theresulting aqueous solution is purified by subjecting it to achromatography through a column packed with Amberlite CG-50 (NH₄ ⁺ form)with the development with water. The eluate fractions containing thedesired compound are collected, concentrated under a reduced pressureand dried, to afford the uridine derivative of the formula (IX) in apure state.

Further, when 5″-N-t-butoxycarbonyl-caprazol as mentioned above istreated in an aqueous solution thereof with methylamine at roomtemperature for a long period of time, it has been found that there canbe produced in its aqueous solution, as a 5″-N-t-butoxycarbonylatedproduct of the uridine derivative of the formula (IX), the compound ofthe following formula (IXa)

The said reaction solution is concentrated under a reduced pressure anddried, to recover the compound of the formula (IXa).

The compound of the formula (IXa) is then dissolved inN,N-dimethylformamide, and to the resulting solution is added an excessamount of an alkylisocyanate of the following general formula (XXIV)

R⁶—NCO  (XXIV)

wherein R⁶ is a straight chain or a substantially straight chain alkylgroup of 1-21 carbon atoms. The reaction intended is then effected atroom temperature. It was thought that by this reaction, there wasproduced a compound which is to be assumed to have the structure of thefollowing general formula (XXV)

wherein R⁶ is the same alkyl group as stated above.

After the reaction between the compound of the formula (IXa) and analkylisocyanate of the formula (XXIV) was effected at room temperaturefor a long period of time, there was deposited a precipitate from theresulting reaction solution. The precipitate was filtered off and thefiltrate was concentrated under a reduced pressure. The resultingconcentrated solution was extracted with chloroform, and the chloroformextract was washed with an aqueous saturated sodium sulfate solution,dried and further concentrated and dried under a reduced pressure. Theresulting solid residue was washed with hexane and dried, to obtain acolorless solid. The colorless solid so obtained was purified by asilica-gel column chromatography (developing withchloroform-water-methanol=9:1:0.1) and then chemically analyzed. Thesolid was recognized to be an imidazolidinone derivative which is aproduct derived from the compound of the estimated formula (XXV) by apartial cyclization thereof, and which is represented by the followinggeneral formula (VIIIa)

wherein R⁶ has the same meaning as defined above.

In order to eliminate the amino-protecting group (Boc) from theimidazolidinone derivative of the formula (VIIIa), the derivative of theformula (VIIIa) was treated with trifluoroacetic acid in methanol. Theresulting reaction solution of the elimination reaction was concentratedto dryness under a reduced pressure, and the resulting residue waswashed with diethyl ether and then dried, to afford an addition salt oftrifluoroacetatic acid of an imidazolidinone derivative of theundermentioned general formula (VIII). This derivative of the formula(VIII) was given CP-IM as code name. The imidazolidinone derivative ofthe formula (VIII) is also found to have antibacterial activitiesagainst bacteria.

According to a tenth aspect of this invention, therefore, there isprovided an imidazolidinone derivative, CP-IM, which is represented bythe following general formula (VIII)

wherein Me is methyl group and R⁶ is a straight chain or a substantiallystraight chain alkyl group of 1-21 carbon atoms, or a pharmaceuticallyacceptable acid addition salt thereof.

Some concrete examples of the derivative of the general formula (VIII)according to the tenth aspect of this invention are shown in thefollowing Table 13 together with their Compound code names and specificrotation data.

TABLE 13 (VIII)

Specific rotation Compound code R⁶ group of the [α]_(D) ²⁰ name formula(VIII) (c 2, in methanol) Compound VIII-A —(CH₂)₅CH₃ Compound VIII-B—(CH₂)₆CH₃ Compound VIII-C —(CH₂)₇CH₃ Compound VIII-D —(CH₂)₈CH₃Compound VIII-E —(CH₂)₉CH₃ +12° Compound VIII-F —(CH₂)₁₀CH₃ +12°Compound VIII-G —(CH₂)₁₁CH₃ +13°

Test Example 6

Minimum growth inhibitory concentrations (mcg/ml) of some of thederivative of the formula (VIII) against a variety of microorganismswere measured on an agar medium by a serial dilution method according tothe standard method as provided by Japanese Society of Chemotherapy. Theresults obtained are shown in the following Table 14.

TABLE 14 Minimum growth inhibitory concentration (mcg/ml) Compound codeagainst bacteria name of the test Staphylococcus MicrococcusStaphylococcus compound (see aureus luteus aureus Table 13) FDA209PFDA16 ATCC607 Compound VIII-A Compound VIII-B Compound VIII-C CompoundVIII-D Compound VIII-E 25 6.25 6.25 Compound VIII-F 25 6.25 6.25Compound VIII-G 25 6.25 12.5

It is to be added that the uridine derivative of the formula (IX) andthe 5″-N-t-butoxycarbonyl-uridine derivative of the formula (IXa) abovedo not have a significant antibacterial activity, but both thederivatives are novel compounds useful as intermediate compounds for usein the synthesis of the derivative of the formula (VIII).

According to an eleventh aspect of this invention, therefore, there isprovided an uridine derivative represented by the following formula (IX)

wherein Me is methyl group, or its 5″-N-t-butoxycarbonyl derivative.

As described hereinbefore, the caprazene-1″′-amide derivative of theformula (II) according to the third aspect of this invention, thecaprazene-1″′-ester derivative of the formula (III) according to thefourth aspect of this invention, the caprazol-1″′-amide derivative ofthe formula (V) according to the seventh aspect of this invention, thecaprazol-1″′-amide-3″′-ester derivative of the formula (VI) according tothe eighth aspect of this invention, the caprazol-3″′-ester derivativeor the caprazol-1″′-ester-3′″-ester derivative of the formula (VII)according to the ninth aspect of this invention or the imidazolidinonederivative, CP-IM, of the formula (VIII) according to the tenth aspectof this invention, or acid addition salts of these derivatives haveantibacterial activities against a variety of bacteria, so that at leastone of these derivatives or their acid addition salts can be used asactive ingredient and can be associated with a pharmaceuticallyacceptable carrier or carriers to form a pharmaceutical or medicinalcomposition, which may be particularly an antibacterial composition.Pharmaceutically acceptable liquid carriers conventionally used may, forexample, include ethanol, aqueous ethanol, water, physiological saltsolution and the like, and solid carriers may, for example, becrystalline cellulose, starch and the like.

The caprazene derivative of the formula (II) or the formula (III), thecaprazol derivative of the formula (V) or the formula (VI) or theformula (VII) or the imidazolidinone derivatives of the formula (VIII)or acid addition salts of these derivatives may be administered, eitherby itself or in the form of a pharmaceutical composition containing itas active ingredient, through any appropriate route.

According to a further aspect of this invention, therefore, there isprovided a pharmaceutical composition comprising as active ingredient atleast one of a caprazene-1″′-amide derivative of the formula (II) or acaprazene-1″′-ester derivative of the formula (III) or acaprazol-1″′-amide derivative of the formula (V) or acaprazol-1″′-amide-3′″-ester derivative of the formula (VI) or acaprazol-3″′-ester derivative or a caprazol-1″′-ester-3″′-esterderivative of the formula (VII) or an imidazolidinone derivative, CP-IM,of the formula (VIII), or an acid addition salt of these derivative, anda pharmaceutically acceptable liquid or solid carrier or carriers, incombination with the active ingredient.

BEST MODE FOR CARRYING OUT THE INVENTION

Now, an illustrative experiment of the preparation of caprazene of theformula (I) according to the first aspect of this invention by theprocess according to the second aspect of this invention is concretelyexplained with reference to the following

Example 1 Synthesis of Caprazene from Caprazamycin B

Caprazamycin B (200 mg) was dissolved in 80% aqueous acetic acidsolution (6 ml) and the resulting solution was heated at 70° C. for 2hours. The resulting reaction solution was concentrated, and to theresulting syrupy concentrate was added an amount of acetone. Theprecipitate so deposited was recovered by filtration, washed withacetone and dried. Thus, there was afforded caprazene (96.3 mg) as acolorless solid. Yield; 99%.

Melting point: 210-211° C. (with decomposition) (after thecrystallization from water-acetone)

Specific rotation: [α]¹⁹+85° (c 0.5, H₂O)

¹H-NMR spectrum and ¹³C-NMR spectrum of caprazene are shown in thefollowing Table 15.

TABLE 15 ¹³C-NMR data of ¹H-NMR data of caprazene caprazene Position (δ,ppm in D₂O) Position (δ, ppm in D₂O) 5 5.82, d, J = 8 Hz 2 151.7 6 7.69,d, J = 8 Hz 4 166.8 5 102.0 6 142.4 1′ 5.62, d, J = 2.5 Hz 2′ 4.28, dd,J = 2.5, 5 Hz 1′ 91.4 3′ 4.12, dd, J = 5, ~8 Hz 2′ 73.9 4′ 4.24, br. d,J = ~8 Hz 3′ 69.4 5′ 4.34, dd, J = 2, 9.5 Hz 4′ 82.7 5′ 77.0 1″ 5.22,slightly br. s 2″ 4.13, br. d, J = ~5 Hz 3″ 4.26, dd, J = ~5, ~8 Hz 1″110.0 4″ 4.20, m 2″ 75.3 5″a 3.18, dd, J = 5, 14 Hz 3″ 70.7 5″b 3.35,dd, J = 4, 14 Hz 4″ 79.0 5″ 40.5 2′″ 3′″ 6.49, t, J = 7 Hz 1′″ 169.24′″a 2.94, dd, J = 7, 12.5 Hz 2′″ 144.7 4′″b 3.34, dd, J = 7, 12.5 Hz3′″ 123.5 6′″ 3.92, d, J = 9.5 Hz 4′″ 51.5 MeN-5′″ 2.42, s 6′″ 63.6(broad) MeN-8′″ 2.99, s 7′″ 171.3 MeN-5′″ 40.5 MeN-8′″ 33.2

Example 2 Synthesis of Caprazene from a Mixture of Caprazamycins B, C,D, E and F

A mixture (10.1 g) of caprazamycins B-F (see the general formulae (A)and (B) shown hereinbefore) was dissolved in 80% aqueous acetic acidsolution (250 ml) and the resulting solution was heated at 70° C. for 2hours. The reaction solution obtained was concentrated, and to theresulting syrupy concentrate was added an amount of acetone, and thedeposited precipitate was recovered by filtration. The solid soprecipitated and recovered was washed with acetone and dried, to affordcaprazene (5.1 g).

Now, an illustrative experiment of the preparation of acaprazene-1″′-amide derivative of the formula (II) according to thethird aspect of this invention is concretely described with reference toExample 3.

Example 3 (a) Synthesis of 5″-N-Boc-Caprazene from Caprazene

Caprazene of the formula (I) (8.14 g) was suspended in a mixed solvent(120 ml) of water-dioxane (2:1). To the resultant suspension was addedtriethylamine (3.7 ml) to give a homogeneous solution of caprazene. Tothe resulting solution was added a solution of di-t-butyl dicarbonate(3.2 g) dissolved in dioxane (5 ml), and the reaction intended wasconducted at room temperature for 1 hour (for the reaction forintroducing t-butoxycarbonyl group (Boc) as amino-protecting group). Thereaction solution obtained was concentrated and the resulting residuewas washed with ethyl acetate and dried, to afford 5″-N-Boc-caprazene(9.50 g) as a pale yellow solid. Crude yield; 99%.

¹H-NMR spectrum (in heavy water (D₂O), TMS internal standard)

δ 1.31 (3H, s, Me₃CO—)

-   -   2.39 (3H, slightly br. s, MeN-5″′)    -   2.98 (3H, s, MeN-8″′)    -   5.13 (1H, slightly br. s, H-1″)    -   5.62 (1H, slightly br. s, H-1′)    -   5.77 (1H, d, H-5, J_(5,6)=8 Hz)    -   6.44 (1H, t, H-3″′, J=7 Hz)    -   5.77 (1H, d, H-6).

(b) Synthesis of Caprazene-1″′-Amide Derivative from 5″-N-Boc-Caprazene

The 5″-N-Boc-caprazene obtained in Example 3(a) (150 mg) was suspendedin tetrahydrofuran (6 ml). To the resulting suspension were addedtriethylamine (80 μl), N,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride(80 mg) and one of the various amine compounds R¹—NH₂ shown in thefollowing Table 16 or one of various para-substituted aniline (1.1 to1.3 molar equivalents each). The resulting mixture was stirred at roomtemperature for 1 hour to cause the reaction intended (for the amidationreaction).

TABLE 16 Amine compound R¹—NH₂ Chemical formula Name C₆H₁₃—NH₂Hexylamine C₇H₁₅—NH₂ Heptylamine C₈H₁₇—NH₂ Octylamine C₉H₁₉—NH₂Nonylamine C₁₀H₂₁—NH₂ Decylamine C₁₁H₂₃—NH₂ Undecylamine C₁₂H₂₅—NH₂Dodecylamine C₁₃H₂₇—NH₂ Tridecylamine C₁₄H₂₉—NH₂ TetradecylamineC₁₅H₃₁—NH₂ Pentadecylamine C₁₆H₃₃—NH₂ Hexadecylamine C₁₇H₃₅—NH₂Heptadecylamine C₁₈H₃₇—NH₂ Octadecylamine C₁₉H₃₉—NH₂ NonadecylamineC₂₀H₄₁—NH₂ Icocylamine C₂₁H₄₃—NH₂ Henicocylamine Cyclo(CH₂)₁₂—NH₂Cyclododecylamine CH₃(CH₂)₇C═C(CH₂)₈—NH₂ Oleylamine

The resulting reaction solution was concentrated and the resultingsyrupy concentrate was extracted with chloroform. The chloroform extractwas washed with water and then concentrated. The resulting residue wasdissolved in chloroform and the chloroform solution was purified by asilica-gel column chromatography (developing solventsystem:chloroform-methanol=10:1). The desired eluate fractions werecollected and concentrated to dryness. Thus, there was afforded 5″-N-Bocprotected derivative of each of the caprazene-1″′-amide derivatives ofthe formula (II) which has Compound code name shown in Table 2 givenhereinbefore, as a colorless solid. Yield; 86-128 mg (the yield in thetwo steps from caprazene; 50-60%).

(c) Synthesis of Caprazene-1″′-Amide Derivative

Each of the 5″-N-Boc protected derivatives of a caprazene-1″′-amidederivative obtained in Example 3 (b) (50 mg) was dissolved in methanolsolution of 80% trifluoroacetic acid (1 ml). The resulting solution wassubjected to the reaction at room temperature for 1 hour to eliminatethe amino-protecting group (Boc). The resulting deprotection reactionsolution was concentrated, and to the resulting syrupy concentrate wasadded an amount of diethyl ether, and the precipitate deposited waswashed with diethyl ether and then dried. Thus, there were affordedcaprazene-1″′-amide derivatives of the formula (II) which are CompoundII-A to Compound II-R shown in Table 2-1 above, or Compound II-1 toCompound II-24 shown in Table 2-2 above, respectively, as a colorlesssolid. Yield; 54.5-58.0 mg (Yield as an addition salt ofbis-trifluoroacetic acid; 96-99%).

¹H-NMR spectrum (500 MHz, in deutero-dimethylsulfoxide, TMS internalstandard) of each of the Compound II-A to Compound II-R (see Table 2-1)or Compound II-1 to Compound II-24 (see Table 2-2) obtained ascaprazene-1″′-amide derivatives of the formula (II) in Example 3(c) isshown below.

Compound II-A

δ 0.84 (3H, t, CH₃ (CH₂)₅NH, J=7 Hz), 1.18-1.25 (6H, slightly br. s, CH₃(CH₂)₃ CH₂CH₂NH), 2.36 (3H, br. s, NMe-5′″), 2.91 (3H, s, NMe-8′″). 5.10(1H, br. s, H-1″), 5.55 (1H, d, H-1′, J=1.5 Hz), 5.63 (1H, d, H-5, J=˜8Hz), 6.31 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. s, H-6), 11.33 (1H,s, NH-3).

Compound II-B

δ 0.84 (3H, t, CH₃ (CH₂)₆NH, J=7 Hz), 1.16-1.28 (8H, br. s, CH₃ (CH₂)₄CH₂CH₂NH), 2.36 (3H, br. s, NMe-5″′), 2.91 (3H, s, NMe-8′″). 5.10 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜1 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.30 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.33(1H, s, NH-3).

Compound II-C

δ 0.85 (3H, t, CH₃ (CH₂)₇NH, J=7 Hz), 1.18-1.28 (10H, br. s, CH₃ (CH₂)₅CH₂CH₂NH), 2.36 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″). 5.09 (1Hbr. s, H-1″), 5.55 (1H, d, H-1′, J=˜1.5 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. s, H-6), 11.32 (1H, s,NH-3).

Compound II-D

δ 0.85 (3H, t, CH₃ (CH₂)₈NH, J=7 Hz), 1.18-1.29 (12H, br. s, CH₃ (CH₂)₆CH₂CH₂NH), 2.34 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.08 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜1 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.68 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-E

δ 0.85 (3H, t, CH₃ (CH₂)₉NH, J=7 Hz), 1.18-1.28 (14H, br. s, CH₃ (CH₂)₇CH₂CH₂NH), 2.34 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜1 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.28 (1H, br. t, H-3″′, J=˜6 Hz), 7.68 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-F

δ 0.85 (3H, t, CH₃ (CH₂)₁₀NH, J=7 Hz), 1.18-1.30 (16H, br. s, CH₃ (CH₂)₈CH₂CH₂NH), 2.36 (3H, br. s, NMe-5″′), 2.91 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜1 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-G

δ 0.85 (3H, t, CH₃ (CH₂)₁₁NH, J=7 Hz), 1.18-1.29 (18H, br. s, CH₃ (CH₂)₉CH₂CH₂NH), 2.34 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.08 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜2 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.68 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-H

δ 0.86 (3H, t, CH₃ (CH₂)₁₂NH, J=7 Hz), 1.18-1.30 (20H, br. s, CH₃ (CH₂)₁₀CH₂CH₂NH), 2.35 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜1 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-I

δ 0.86 (3H, t, CH₃ (CH₂)₁₃NH, J=7 Hz), 1.18-1.30 (22H, br. s, CH₃ (CH₂)₁₁CH₂CH₂NH), 2.35 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, s, H-1′), 5.63 (1H, d, H-5, J=˜8 Hz), 6.29 (1H,br. t, H-3″′, J=˜6 Hz), 7.68 (1H, br. d, H-6, J=˜8 Hz), 11.32 (1H, s,NH-3).

Compound II-J

δ 0.86 (3H, t, CH₃ (CH₂)₁₄NH, J=7 Hz), 1.18-1.30 (24H, br. s, CH₃ (CH₂)₁₂CH₂CH₂NH), 2.35 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, s, H-1′), 5.63 (1H, d, H-5, J=˜8 Hz), 6.29 (1H,br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.32 (1H, s,NH-3).

Compound II-K

δ 0.85 (3H, t, CH₃ (CH₂)₁₅NH, J=7 Hz), 1.18-1.30 (26H, br. s, CH₃ (CH₂)₁₃CH₂CH₂NH), 2.36 (3H, br. s, NMe-5″′), 2.91 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜2 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-L

δ 0.85 (3H, t, CH₃ (CH₂)₁₆NH, J=7 Hz), 1.18-1.30 (28H, br. s, CH₃ (CH₂)₁₄CH₂CH₂NH), 2.35 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, s, H-1′), 5.63 (1H, d, H-5, J=˜8 Hz), 6.29 (1H,br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.32 (1H, s,NH-3).

Compound II-M

δ 0.86 (3H, t, CH₃ (CH₂)₁₇NH, J=7 Hz), 1.18-1.30 (30H, br. s, CH₃ (CH₂)₁₅CH₂CH₂NH), 2.35 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.09 (1H,br. s, H-1″), 5.55 (1H, s, H-1′), 5.63 (1H, d, H-5, J=˜8 Hz), 6.29 (1H,br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6, J=˜8 Hz), 11.32 (1H, s,NH-3).

Compound II-N

δ 0.85 (3H, t, CH₃ (CH₂)₁₈NH, J=7 Hz), 1.18-1.30 (32H, br. s, CH₃ (CH₂)₁₆CH₂CH₂NH), 2.34 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.08 (1H,br. s, H-1″), 5.55 (1H, d, H-1′, J=˜2 Hz), 5.63 (1H, d, H-5, J=˜8 Hz),6.29 (1H, br. t, H-3″′, J=˜6 Hz), 7.68 (1H, br. d, H-6, J=˜8 Hz), 11.32(1H, s, NH-3).

Compound II-O

δ 0.85 (3H, t, CH₃ (CH₂)₁₉NH, J=7 Hz), 1.18-1.30 (34H, br. s, CH₃ (CH₂)₁₇CH₂CH₂NH), 2.34 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.08 (1H,br. s, H-1″), 5.55 (1H, slightly br. d, H-1′, J=˜1 Hz), 5.63 (1H, d,H-5, J=˜8 Hz), 6.28 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br. d, H-6,J=˜8 Hz), 11.32 (1H, s, NH-3).

Compound II-P

δ 0.86 (3H, t, CH₃ (CH₂)₂₀NH, J=7 Hz), 1.18-1.30 (36H, br. s, CH₃ (CH₂)₁₈CH₂CH₂NH), 2.34 (3H, br. s, NMe-5″′), 2.90 (3H, s, NMe-8′″), 5.08 (1H,br. s, H-1″), 5.55 (1H, slightly br. d, H-1′, J=˜1 Hz), 5.63 (1H, d,H-5, J=˜8 Hz), 6.28 (1H, br. t, H-3″′, J=˜6 Hz), 7.68 (1H, br. d, H-6,J=˜8 Hz), 11.31 (1H, s, NH-3).

Compound II-Q

δ 1.14-1.45 (22H, m, —(CH₂) ₁₁—), 2.35 (3H, br. s, NMe-5″′), 2.91 (3H,s, NMe-8″′), 5.09 (1H, br. s, H-1″), 5.58 (1H, d, H-1′, J=˜2 Hz), 5.64(1H, d, H-5, J=˜8 Hz), 6.31 (1H, br. t, H-3″′, J=˜6 Hz), 7.66 (1H, br.d, H-6, J=˜8 Hz), 11.33 (1H, s, NH-3).

Compound II-R

δ 0.85 (3H, t, CH₃ CH₂—, J=7 Hz), 2.35 (3H, br. s, NMe-5″′), 2.90 (3H,s, NMe-8″′), 5.09 (1H, br. s, H-1″), 5.55 (1H, d, H-1′, J=˜2 Hz), 5.63(1H, d, H-5, J=˜8 Hz), 6.28 (1H, br. t, H-3″′, J=˜6 Hz), 7.67 (1H, br.d, H-6, J=˜8 Hz), 11.32 (1H, s, NH-3).

Compound II-1

δ 2.26 (3H, s, CH₃ C₆H₄NH), 2.38 (3H, br. s, NMe-5″′), 2.96 (3H, s,NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2 Hz), 5.62 (1H,d, H-5, J=˜8 Hz), 6.40 (1H, br. t, H-3″′, J=˜6 Hz), 7.13 and 7.50 (each2H, d, CH₃ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=˜8 Hz), 10.14 (1H, s,CH₃C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-2

δ 1.16 (3H, t, CH₃ CH₂C₆H₄NH, J=8 Hz), 2.37 (3H, br. s, NMe-5′″), 2.95(3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2 Hz),5.62 (1H, dd, H-5, J=2, 8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.15and 7.52 (each 2H, d, CH₃CH₂ C₆H₄ NH, J=8 Hz), 7.69 (1H, d, H-6, J=8Hz), 10.14 (1H, s, CH₃CH₂C₆H₄ NH), 11.32 (1H, d, NH-3, J=2 Hz).

Compound II-3

δ 0.87 (3H, t, CH₃ (CH₂)₂C₆H₄NH, J=8 Hz), 2.37 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, dd, H-5, J=2, 8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz),7.14 and 7.52 (each 2H, d, CH₃(CH₂)₂ C₆H₄ NH, J=8 Hz), 7.69 (1H, d, H-6,J=8 Hz), 10.14 (1H, s, CH₃(CH₂)₂C₆H₄ NH), 11.32 (1H, d, NH-3, J=2 Hz).

Compound II-4

δ 0.89 (3H, t, CH₃ (CH₂)₃C₆H₄NH, J=7.5 Hz), 2.38 (3H, br. s, NMe-5″′),2.96 (3H, s, NMe-8″′), 5.12 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.40 (1H, br. t, H-3″′, J=˜6 Hz), 7.14and 7.52 (each 2H, d, CH₃(CH₂)₃ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.15 (1H, s, CH₃(CH₂)₃C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-5

δ 0.85 (3H, t, CH₃ (CH₂)₄C₆H₄NH, J=7 Hz), 2.38 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.13and 7.51 (each 2H, d, CH₃(CH₂)₄ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.14 (1H, s, CH₃(CH₂)₄C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-6

δ 0.85 (3H, t, CH₃ (CH₂)₅C₆H₄NH, J=7 Hz), 2.38 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.13and 7.51 (each 2H, d, CH₃(CH₂)₅ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.14 (1H, s, CH₃(CH₂)₅C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-7

δ 0.85 (3H, t, CH₃ (CH₂)₆C₆H₄NH, J=7 Hz), 2.37 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.13and 7.51 (each 2H, d, CH₃(CH₂)₆ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.14 (1H, s, CH₃(CH₂)₆C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-8

δ 0.85 (3H, t, CH₃ (CH₂)₂C₆H₄NH, J=7 Hz), 2.37 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.12 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.13and 7.51 (each 2H, d, CH₃(CH₂)₇ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.14 (1H, s, CH₃(CH₂)₂C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-10

δ 0.85 (3H, t, CH₃ (CH₂)₉C₆H₄NH, J=7 Hz), 2.38 (3H, br. s, NMe-5″′),2.96 (3H, s, NMe-8″′), 5.12 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.13and 7.51 (each 2H, d, CH₃(CH₂)₉ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.14 (1H, s, CH₃(CH₂)₉C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-12

δ 0.85 (3H, t, CH₃ (CH₂)₁₁C₆H₄NH, J=8 Hz), 2.38 (3H, br. s, NMe-5″′),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=˜2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 7.13and 7.51 (each 2H, d, CH₃(CH₂)₁₁ C₆H₄ NH, J=8 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.15 (1H, s, CH₃(CH₂)₁₁C₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-14

δ 0.85 (3H, t, CH₃ (CH₂)₁₃C₆H₄NH, J=7 Hz), 2.37 (3H, br. s, NMe-5″′),2.95 (3H, s, NMe-8″′), 5.12 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, dd, H-5, J=2, ˜8 Hz), 6.38 (1H, br. t, H-3″′, J=˜6 Hz),7.13 and 7.51 (each 2H, d, CH₃(CH₂)₁₃ C₆H₄ NH, J=8.5 Hz), 7.69 (1H, d,H-6, J=8 Hz), 10.14 (1H, s, CH₃(CH₂)₁₃C₆H₄ NH), 11.31 (1H, d, NH-3, J=˜2Hz).

Compound II-15

δ 2.37 (3H, br. s, NMe-5″′), 2.95 (3H, s, NMe-8″′), 3.73 (3H, s, OCH₃),5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2 Hz), 5.63 (1H, dd, H-5,J=˜2, 8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 6.89 and 7.52 (each 2H,d, CH₃OC₆H₄ NH, J=9 Hz), 7.68 (1H, d, H-6, J=8 Hz), 10.08 (1H, s,CH₃OC₆H₄ NH), 11.32 (1H, d, NH-3, J=˜2 Hz).

Compound II-16

δ 1.31 (3H, t, CH₃ CH₂OC₆H₄NH, J=7 Hz), 2.38 (3H, br. s, NMe-5″′), 2.96(3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2 Hz),5.63 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 6.87 and7.51 (each 2H, d, CH₃CH₂OC₆H₄ NH, J=9 Hz), 7.68 (1H, d, H-6, J=8 Hz),10.08 (1H, s, CH₃CH₂OC₆H₄ NH), 11.33 (1H, s, NH-3).

Compound II-18

δ 0.93 (3H, t, CH₃ (CH₂)₃OC₆H₄NH, J=7.5 Hz), 2.37 (3H, br. s, NMe-5″′),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 6.88and 7.51 (each 2H, d, CH₃(CH₂)₃OC₆H₄ NH, J=9 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.07 (1H, s, CH₃(CH₂)₃OC₆H₄ NH), 11.33 (1H, s, NH-3).

Compound II-19

δ 0.89 (3H, t, CH₃ (CH₂)₄OC₆H₄NH, J=7 Hz), 2.37 (3H, br. s, NMe-5′″),2.96 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 6.88and 7.50 (each 2H, d, CH₃(CH₂)₄OC₆H₄ NH, J=9 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.07 (1H, s, CH₃(CH₂)₄OC₆H₄ NH), 11.33 (1H, s, NH-3).

Compound II-20

δ 0.88 (3H, t, CH₃ (CH₂)₅OC₆H₄NH, J=7 Hz), 2.38 (3H, br. s, NMe-5′″),2.96 (3H, s, NMe-8″′), 5.12 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 6.88and 7.50 (each 2H, d, CH₃(CH₂)₅OC₆H₄ NH, J=9 Hz), 7.67 (1H, d, H-6, J=8Hz), 10.08 (1H, s, CH₃(CH₂)₅OC₆H₄ NH), 11.33 (1H, s, NH-3).

Compound II-21

δ 0.87 (3H, t, CH₃ (CH₂)₆OC₆H₄NH, J=7 Hz), 2.37 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.39 (1H, br. t, H-3″′, J=˜6 Hz), 6.88and 7.50 (each 2H, d, CH₃(CH₂)₆OC₆H₄ NH, J=9 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.07 (1H, s, CH₃(CH₂)₆OC₆H₄ NH), 11.32 (1H, s, NH-3).

Compound II-23

δ 0.86 (3H, t, CH₃ (CH₂)₈OC₆H₄NH, J=7 Hz), 2.37 (3H, br. s, NMe-5′″),2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s, H-1″), 5.60 (1H, d, H-1′, J=2Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.38 (1H, br. t, H-3″′, J=˜6 Hz), 6.88and 7.50 (each 2H, d, CH₃(CH₂)₈OC₆H₄ NH, J=9 Hz), 7.68 (1H, d, H-6, J=8Hz), 10.07 (1H, s, CH₃(CH₂)₈OC₆H₄ NH), 11.32 (1H, d, NH-3, J=˜2 Hz).

Compound II-24

δ 2.37 (3H, br. s, NMe-5″′), 2.95 (3H, s, NMe-8″′), 5.11 (1H, br. s,H-1″), 5.59 (1H, d, H-1′, J=2 Hz), 5.62 (1H, d, H-5, J=˜8 Hz), 6.38 (1H,br. t, H-3″′, J=˜6 Hz), 7.16 and 7.52 (each 2H, d, —C₆H₄ NH, J=9 Hz),7.68 (1H, d, H-6, J=8 Hz), 10.14 (1H, s, —C₆H₄ NH), 11.32 (1H, s, NH-3).

Now, an illustrative experiment of the preparation of acaprazene-1″′-ester derivative of the formula (III) according to thefourth aspect of this invention is concretely described with referenceto the following Example 4.

Example 4 (a) Preparation of 5″-N-Boc-caprazene-1″′-ester derivativefrom 5″-N-Boc-caprazene

The 5″-N-Boc-caprazene obtained in Example 3(a) (150 mg) was dissolvedin pyridine (5 ml). To the resultant solution were addedN,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride (120 mg) as well aseach one of a variety of alcohol compounds R²—OH shown in Table 17 below(2 molar equivalents in each case). The resulting reaction mixture wasstirred at room temperature overnight to cause the reaction intended(for the esterification reaction).

TABLE 17 Alcohol compound R²—OH Chemical formula Name H₃C(CH₂)₉—OH Decylalcohol H₃C(CH₂)₁₂—OH Tridecyl alcohol H₃C(CH₂)₁₇—OH Octadecyl alcoholH₃C—CH₂—CH═CH—(CH₂)₁₀—OH Cis-11-tetradecene-1-ol H₃C—(CH₂)₈—CH═CH—CH₂—OHTrans-2-dodecenol H₂C═CH—(CH₂)₉—OH 10-Undecene-1-olH₃C—(CH₂)₅—C≡C—(CH₂)₂—OH 3-Decyne-1-ol

The resulting esterification reaction solution was concentrated and theresulting syrupy concentrate was extracted with chloroform. Thechloroform extract was washed with water and then concentrated. Theresulting residue was purified by a silica-gel column chromatography(developing solvent system:chloroform-methanol=10:1). Thus, there wasyielded each of 5″-N-Boc-protected derivatives of caprazene-1″′-esterderivatives having Compound code name shown in Table 5 givenhereinbefore, as a colorless solid. Yield; 96-108 mg (Yield at the endof the two steps from caprazene; 45-52%).

(b) Synthesis of caprazene-1″′-ester derivative

Each of the 5″-N-Boc-caprazene-1″′-ester derivatives obtained in Example4 (a) (50 mg) was dissolved in methanolic solution of 80%trifluoroacetic acid (1 ml). The resultant solution was kept at roomtemperature for 1 hour to cause the reaction for the elimination of theamino-protecting group (Boc). The resulting reaction solution wasconcentrated, and to the resulting syrupy concentrate was added anamount of diethyl ether to deposit a precipitate which was then washedwith diethyl ether and dried. Thus, there was yielded each of CompoundIII-AA to Compound III-GG shown as code name in Table 5 givenhereinbefore, a as colorless solid. Yield; 55.9-57.4 mg (Yield asaddition salt of bis-trifluoroacetic acid; 98-99%).

¹H-NMR spectrum (500 MHz, in deutero-dimethylsulfoxide, TMS internalstandard) of each of the Compound III-AA to Compound III-GG (see Table5) obtained as the caprazene-1″′-ester derivatives of the formula (III)in Example 4(c) is shown below.

Compound III-AA

δ 0.86 (3H, t, CH₃ (CH₂)₉O, J=7 Hz), 1.18-1.35 (14H, slightly br. s, CH₃(CH₂) ₇CH₂CH₂O), 2.35 (3H, br. s, NMe-5″′), 2.96 (3H, s, NMe-8′″), 5.09(1H, s, H-1″), 5.53 (1H, d, H-1′, J=2.5 Hz), 5.64 (1H, dd, H-5, J=˜1.5,8 Hz), 6.73 (1H, t, H-3″′, J=7 Hz), 7.63 (1H, br. d, H-6, J=8 Hz), 11.33(1H, d, NH-3, J=˜1.5 Hz).

Compound III-BB

δ 0.86 (3H, t, CH₃ (CH₂)₁₂O, J=7 Hz), 1.2-1.3 (20H, slightly br. s, CH₃(CH₂) ₁₀CH₂CH₂O), 2.36 (3H, br. s, NMe-5″′), 2.96 (3H, s, NMe-8′″), 5.09(1H, s, H-1″), 5.53 (1H, d, H-1′, J=2 Hz), 5.64 (1H, d, H-5, J=8 Hz),6.73 (1H, t, H-3″′, J=7 Hz), 7.63 (1H, d, H-6, J=8 Hz), 11.33 (1H, s,NH-3).

Compound III-CC

δ 0.86 (3H, t, CH₃ (CH₂)₁₇O, J=7 Hz), 1.2-1.3 (30H, slightly br. s, CH₃(CH₂) ₁₅CH₂CH₂O), 2.37 (3H, br. s, NMe-5″′), 2.97 (3H, s, NMe-8′″), 5.10(1H, slightly br. s, H-1″), 5.53 (1H, d, H-1′, J=2.5 Hz), 5.64 (1H,slightly br. d, H-5, J=8 Hz), 6.74 (1H, t, H-3″′, J=7 Hz), 7.62 (1H, d,H-6, J=8 Hz), 11.33 (1H, slightly br. s, NH-3).

Compound III-DD

δ 0.91 (3H, t, CH₃ CH₂CH═CH—, J=7.5 Hz), 2.36 (3H, br. s, NMe-5′″), 2.96(3H, s, NMe-8″′), 5.09 (1H, s, H-1″), 5.53 (1H, d, H-1′, J=2 Hz), 5.64(1H, d, H-5, J=8 Hz), 6.73 (1H, t, H-3″′, J=7 Hz), 7.63 (1H, d, H-6, J=8Hz), 11.33 (1H, s, NH-3).

Compound III-EE

δ 0.86 (3H, t, CH₃ CH₂—, J=7 Hz), 2.36 (3H, slightly br. s, NMe-5′″),2.96 (3H, s, NMe-8′″), 4.60 (2H, m, —CH₂CH═CHCH₂ O—), 5.09 (1H, s,H-1″), 5.53 (1H, s, H-1′), 5.55 (1H, m, —CH₂CH═CHCH₂O—), 5.65 (1H, d,H-5, J=8 Hz), 5.80 (1H, dt, —CH₂ CH═CHCH₂O—, J=˜7, ˜7, ˜15 Hz), 6.75(1H, t, H-3″′, J=7 Hz), 7.64 (1H, d, H-6, J=8 Hz), 11.34 (1H, s, NH-3).

Compound III-FF

δ 2.36 (3H, br. s, NMe-5″′), 2.96 (3H, s, NMe-8″′), 5.09 (1H, br. s,H-1″), 5.53 (1H, slightly br. s, H-1′), 5.65 (1H, d, H-5, J=8 Hz), 5.75(1H, m, CH₂═CH—), 6.73 (1H, t, H-3″′, J=7 Hz), 7.63 (1H, d, H-6, J=8Hz), 11.34 (1H, s, NH-3).

Compound III-GG

δ 0.85 (3H, t, CH₃ CH₂—, J=7 Hz), 2.35 (3H, slightly br. s, NMe-5′″),2.99 (3H, s, NMe-8″′), 5.10 (1H, s, H-1″), 5.55 (1H, d, H-1′, J=˜2 Hz),5.65 (1H, dd, H-5, J=˜1.5, 8 Hz), 6.76 (1H, t, H-3″′, J=7 Hz), 7.65 (1H,d, H-6, J=8 Hz), 11.34 (1H, slightly br. s, NH-3).

Further, an illustrative experiment of the preparation of caprazol ofthe formula (IV) according to the fifth aspect of this invention by theprocess according to the sixth aspect of this invention is concretelyexplained with reference to the following Examples 5-6.

Example 5 Synthesis of Caprazol from Caprazamycin B

Caprazamycin B (150 mg) was dissolved in N,N-dimethylformamide (1.5 ml),and to the resultant solution was then added 28% aqueous ammoniasolution (1.5 ml). The resulting mixture was stirred at room temperaturefor 4 days to effect the intended hydrolysis. Insolubles formed in thereaction solution were filtered off, and thereafter the reactionsolution was concentrated and the resulting residue was washed withacetone and dried. Thus, there was afforded caprazol (74.7 mg) as acolorless solid.

Yield; 99%.

Melting point: 205-206° C. (with decomposition) (after thecrystallization from water-methanol)

Specific rotation: [α]_(D) ¹⁹+28° (c 0.5, dimethylsulfoxide)

¹H-NMR spectrum and ¹³C-NMR spectrum of caprazol are shown in thefollowing Table 18.

TABLE 18 ¹³C-NMR data of ¹H-NMR data of caprazol caprazol Position (δ,ppm in D₂O) Position (δ, ppm in D₂O) 5 5.82, d, J = 8 Hz 2 151.8 6 7.77,d, J = 8 Hz 4 167.1 5 101.7 6 142.9 1′ 5.60, slightly br. s 2′ 4.31, br.d, J = 5 Hz 1′ 91.8 3′ 4.08, dd, J = 5, 8 Hz 2′ 74.0 4′ 4.13, d, J = ~ 8Hz 3′ 69.3 5′ 4.39, d, J = 9 Hz 4′ 82.4 5′ 77.6 1″ 5.17, slightly br. s2″ 4.14, d, J = ~ 3 Hz 3″ 4.25, m 1″ 111.2 4″ ~4.21, m 2″ 75.4 5″a 3.20,dd, J = 4, 13.5 Hz 3″ 70.6 5″b 3.32, dd, J = 3.5, 13.5 Hz 4″ 79.0 5″40.2 2′″ 4.20, d, J = ~ 5 Hz 3′″ 4.44, br. s 1′″ 174.1 4′″a 3.01, br. d,J = 15 Hz 2′″ 70.0 4′″b 3.13, br. d, J = 15 Hz 3′″ 69.3 6′″ 3.85, d, J =9 Hz 4′″ 59.1 MeN-5′″ 2.43, s 6′″ 63.5 MeN-8′″ 3.07, s 7′″ 172.7 MeN-5′″37.0 MeN-8′″ 39.2

Example 6 Synthesis of Caprazol from a Mixture of Caprazamycins C-F

A mixture of caprazamycins C, D, E and F (2.1 g) was dissolved inN,N-dimethylformamide (20 ml). To the resulting solution was added 28%aqueous ammonia solution (20 ml), and then the intended hydrolysisreaction was effected at room temperature for 110 hours. Insolublesformed in the reaction solution was filtered off. Thereafter, thereaction solution was concentrated and the resulting residue was washedwith acetone and then dried. Thus, there was yielded caprazol (1.08 g).

Example 7 Preparation of Caprazene from Caprazol

An amount of caprazol was dissolved in an amount of 1N aqueoushydrochloric acid and the resultant solution was heated at 100° C. for 3hours, thereby to produce caprazene in the yield of 20%. About 20% ofcaprazol was left unreacted. Respective chemical structure of eachcompound was confirmed by NMR analysis. The resulting reaction solutionwas concentrated under a reduced pressure and the resulting residue wassubjected to a silica-gel column chromatography. Thus, there can beseparated caprazene and caprazol, from each other.

Further, an illustrative experiment of the preparation of acaprazol-1″′-amide derivative of the formula (V) according to theseventh aspect of this invention is concretely described with referenceto Example 8 given below.

Example 8 (a) Synthesis of 5″-N-Boc-caprazol

Caprazol of the formula (IV) (2.80 g) was dissolved in a mixed solventof water-dioxane (1:2), and to the resultant solution were addedtriethylamine (1.7 ml) and a dioxane solution (5 ml) of di-t-butyldicarbonate (1.27 g). The intended reaction for the resulting mixturewas effected at room temperature for 1 hour. The resulting reactionsolution was concentrated and the residue obtained was washed with ethylacetate and then dried. Thus, there was afforded 5″-N-Boc-caprazol (3.19g) as a pale yellow solid. Crude yield; 97%.

¹H-NMR spectrum (500 MHz, in D₂O):

δ 1.40 (9H, s, methyl in the t-butoxycarbonyl group), 2.47 (3H, s,NMe-5″′), 3.13 (3H, s, NMe-8″′), 5.16 (1H, s, H-1″), 5.74 (1H, br. s,H-1′).

(b) Synthesis of 1″′-dodecylamide derivative of 5″-N-Boc-caprazol

The 5″-N-Boc-caprazol (97.8 mg) obtained in Example 8(a) was dissolvedin N,N-dimethylformamide (3 ml), and to the resultant solution wereadded triethylamine (0.21 ml), n-dodecylamine (137 mg) andN,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride (188 mg). The resultingmixture was heated at 40° C. to effect the reaction intended (for theamidation reaction). At the end of 2 hours and 4 hours after the startof the reaction, respectively, there were added triethylamine (0.21 ml),n-dodecylamine (137 mg) and N,N-bis(2-oxo-3-oxazolidinyl)phosphinicchloride (189 mg) to the reaction mixture.

At the six hours after the start of the reaction, the reaction solutionwas concentrated to dryness. The residue was extracted with chloroform,and the chloroform extract was washed once with water, then twice withaqueous saturated sodium sulfate solution and then dried over anhydroussodium sulfate. The resulting solution was concentrated to dryness toyield a solid. The solid was purified by a silica-gel columnchromatography (developing system:chloroform-methanol-concentratedaqueous ammonia; 4:1:0.1). Thus there was afforded5″-N-Boc-caprazol-1″′-dodecylamide derivative (45.4 mg) (Yield fromcaprazol; 32%).

(c) Synthesis of caprazol-1″′-dodecylamide derivative

The 5″-N-Boc-caprazol-1″′-dodecylamide derivative above was dissolved ina mixture of trifluoroacetic acid-methanol (8:2) (0.45 ml), and thereaction intended was effected at room temperature for 2 hours (for theelimination of Boc). The reaction solution so obtained was concentratedto dryness, and to the resulting residue was added diethyl ether and theinsoluble matters were washed with diethyl ether. Thus, there wasafforded caprazol-1″′-dodecyl-amide derivative (corresponding toCompound V-G in Table 7) in the form of an addition salt ofbis-trifluoroacetic acid (46 mg) (Yield; 33%).

[α]_(D) ¹⁹+12° (c 1, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol):

δ 0.89 (3H, t, N(CH₂)₁₁ Me, J=7 Hz), 2.51 (3H, s, NMe-5″′), 3.18 (3H, s,NMe-8″′), 5.17 (1H, s, H-1″), 5.76 (1H, s, H-1′), 5.77 (1H, d, H-5,J_(5,6)=8 Hz), 8.08 (1H, d, H-6).

Now, an illustrative experiment of the preparation of acaprazol-1″′-amide-1″′-ester derivative of the formula (VI) according tothe eighth aspect of this invention is explained with reference toExample 9

Example 9 (a) Synthesis of5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol (the compound of theformula (XIV) above)

The 5″-N-Boc-caprazol obtained in Example 8(a) (1.097 g) was dissolvedin N,N-dimethylformamide (33 ml), and the resultant solution were added(±)10-camphor-sulfonic acid (1.06 g) and dimethoxymethane (6 ml) and thereaction intend was effected at room temperature. One day later, therewere further added (±) 10-camphor-sulfonic acid (151 mg) anddimethoxymethane (2 ml). Two days later, there were added (±)10-camphor-sulfonic acid (113 mg) (as acid catalyst) anddimethoxymethane (2 ml). Three days later, there was addeddimethoxymethane (2 ml) to the resulting mixture (for the reaction ofintroducing O-isopropylidene groups).

The 5″-N-Boc-caprazol (3.19 g) obtained in Example 8(a) was dissolved inN,N-dimethylformamide (60 ml), and to the resultant solution were added(±)camphor-10-sulfonic acid (3.29 g) and 2,2-dimethoxypropane (17 ml).The reaction intended was effected at room temperature overnight (forthe reaction of introducing O-isopropylidene groups).

The reaction solution so obtained was neutralized with the addition ofconcentrated aqueous ammonia (0.5 ml) and the neutralized solution wasconcentrated. The resulting residue was dissolved in n-butanol, and theorganic layer was washed with water, then concentrated under a reducedpressure and dried, affording the compound of the formula (XIV) above(3.55 g).

Yield; 99%.

[α]_(D) ¹⁹−30° (c 2, chloroform)

¹H-NMR spectrum (500 MHz, in deutero-methanol):

δ 1.25, 1.36, 1.40, 1.53 (each 3H, s, methyl in isopropylidene group),1.45 (9H, s, methyl in t-butoxycarbonyl group), 2.50 (3H, s, NMe-5″′),3.09 (3H, s, NMe-8″′), 5.24 (1H, s, H-1″), 5.83 (1H, d, H-1′,J_(1′,2′)=2.7 Hz).

(b) Preparation of5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-dodecylamidederivative (a compound included within the derivative of the formula(XV) above)

The 5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol (69.6 mg) obtainedin Example 9(a) was dissolved in N,N-dimethylformamide (1.78 ml), and tothe resultant solution were added triethylamine (0.052 ml),n-dodecylamine (34.2 mg) and N,N-bis(2-oxo-3-oxazolidinyl)phosphinicchloride (47 mg). Then, reaction intended was effected at roomtemperature (for the 1″′-amidation reaction). At the end of two hoursafter and four hours after the start of the reaction, respectively,there were further added triethylamine (0.052 ml), n-dodecylamine (34.2mg) and N,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride (47 mg).

After the reaction was effected for 8 hours, the reaction solutionobtained was concentrated to dryness and the residue was extracted withchloroform. The chloroform extract was washed with water, dried overanhydrous sodium sulfate and concentrated to dryness. The resultingresidue was purified by a silica-gel column chromatography (developingsystem:chloroform-methanol, 50:1). Thus, there was afforded5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-dodecylamidederivative (45.8 mg)

(Yield; 54%).

[α]_(D) ¹⁹−31° (c 2, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol):

δ 0.89 (3H, t, N(CH₂)₁₁ Me, J=7 Hz), 1.44 (9H, s, methyl int-butoxycarbonyl group), 2.50 (3H, s, NMe-5″′), 3.16 (3H, s, NMe-8″′),5.27 (1H, s, H-1″), 5.73 (1H, d, H-5, J_(5,6)=8 Hz), 5.94 (1H, d, H-1′,J_(1′,2′)=2.7 Hz), 7.72 (1H, d, H-6).

(c) Synthesis of5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-dodecylamide-3″′-dodecanoyl-esterderivative (a compound included within the derivative of the formula(XVII) above)

The 5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-dodecylamidederivative (45.5 mg) obtained in Example 9(b) was dissolved indichloromethane. To the resulting solution, under ice-cooling, wereadded 4-dimethylaminopyridine (24.5 mg) and dodecanoyl chloride (0.035ml) (trivial name: lauroyl chloride, Cl—CO—(CH₂)₁₀—CH₃) as acylatingreagent. And the resulting mixture was subjected to the reactionintended under ice-cooling for 3 hours (for the 3″′-O-esterification).Methanol (0.027 ml) was added to the resulting reaction solution, andthen the resulting solution was diluted with chloroform. The resultingmixed solution was washed with aqueous 10% potassium hydrogen sulfatesolution and then with water, dried over anhydrous sodium sulfate andthen concentrated to dryness. The residue so obtained was purified by asilica-gel column chromatography (developing solventsystem:chloroform-methanol, 150:0→150:1), thus affording the titledcompound (39.2 mg; yield; 72%).

(d) Synthesis of caprazol-1″′-dodecylamide-3″′-dodecanoyl-esterderivative (compound corresponding to Compound VI-G in Table 9 includedwithin the eighth aspect of this invention)

The compound as obtained in the above step (c) was dissolved in amixture of trifluoroacetic acid-methanol (8:2) (0.35 ml), and theresulting solution was subjected to the reaction intended at roomtemperature for 4 hours (for the deprotecting reaction). The reactionsolution obtained was concentrated to dryness, and to the resultantresidue was added diethyl ether. The resulting insolubles were washedwith diethyl ether. Thus, there was afforded the titled compound, i.e.caprazol-1″′-dodecylamide-3″′-dodecanoylester derivative (Compound VI-G)in the form of an addition salt of bis-trifluoroacetic acid (35.8 mg;yield from the compound of the step(c) above; 63%).

[α]_(D) ²¹+6° (c 0.5, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol):

δ 0.90 (6H, t, N(CH₂)₁₁ Me and (CH₂)₁₀ Me, J=7 Hz), 2.37 (2H, t,CH₂(CH₂)₉Me, J=7 Hz), 2.45 (3H, s, NMe-5″′), 3.16 (3H, s, NMe-8″′), 5.18(1H, s, H-1″), 5.53 (1H, br. s, 7.73 (1H, d, H-6, J_(5,6)=8 Hz).

Further, an illustrative experiment of the preparation ofcaprazol-3″′-ester derivative of the formula (VII) according to theninth aspect of this invention is explained with reference to thefollowing Example 10.

Example 10 (a) Synthesis of 5″-N-Boc-2′,3′;2″,3″-di-O—isopropylidene-caprazol-3″′-dodecanoyl-ester derivative (a compoundincluded within the derivatives of the formula (XVIII) above)

5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol, i.e. the compound ofthe formula (XIV) above, as obtained in Example 9(a) (42 mg) wasdissolved in dichloromethane (0.84 ml). To the resultant solution, underice-cooling, were added 4-dimethylaminopyridine (13.6 mg) and dodecanoylchloride (0.019 ml) [Cl—CO—(CH₂)₁₀—CH₃; one of acid chlorides of theformula (XVI)]. The resulting mixture was subjected to the reactionintended under ice-cooling (for the 3″′-O-esterification). After the endof 7 hours of the reaction, there were further added4-dimethylaminopyridine (13.6 mg) and dodecanoyl chloride (0.019 ml).After the 24 hours of the reaction, there were added4-dimethylaminopyridine (11.2 mg) and dodecanoyl chloride (0.019 ml),and after the 36 hours of the reaction, there were further added4-dimethylaminopyridine (11.9 mg) and dodecanoyl chloride (0.019 ml) andthe reaction was proceeded further.

After the 48 hours of the reaction, the resulting esterifying reactionsolution, after the addition of methanol (0.017 ml) thereto, was dilutedwith chloroform. The resulting mixture was washed with 10% aqueouspotassium hydrogen sulfate solution and then with water and then driedover anhydrous sodium sulfate. The resulting dried solution wasconcentrated to dryness, to afford a solid containing the titledcompound (82.7 mg).

(b) Synthesis of caprazol-3″′-dodecanoyl ester derivative of thefollowing formula (VII-1) (which is the compound included within thederivatives of the formula (XIX) or the formula (VII) above and whichcorresponds to Compound VII-T in Table 11)

The solid containing the N,O-protected caprazol-3″′-dodecanoyl-esterderivative as obtained in the step(a) above (82.7 mg) was dissolved in amixture of trifluoroacetic acid-methanol (8:2) (0.85 ml). The resultantsolution was subjected to the reaction intended at room temperature for2.5 hours (for the deprotecting reaction). The resulting reactionsolution was concentrated to dryness, and to the resultant residue wasadded diethyl ether. The resulting insolubles were washed with diethylether to give a solid (28.8 mg). The resulting solid was suspended inwater and the suspension was passed through a column packed with Diaion(Registered Trade Mark) HP-20. After the column was washed with water,the column was eluted with 50% aqueous methanol, 80% aqueous methanol,and methanol, in order. The eluate fractions containing the desiredsubstance were concentrated to dryness, thus affording the titledcompound (10.4 mg) (yield from the compound obtained in the step (a)above; 25%).

[α]_(D) ²⁰+17° (c 0.5, dimethylsulfoxide)

¹H-NMR spectrum (500 MHz, in deutero-dimethylsulfoxide):

δ 0.86 (3H, t, (CH₂)₁₀ Me, J=7 Hz), 2.26 (3H, s, NMe-5″′), 2.93 (3H, s,NMe-8″′), 5.00 (1H, s, H-1″), 5.40 (1H, br. s, H-3″′), 5.56 (1H, s,H-1′), 5.64 (1H, d, H-5, J_(5,6)=8 Hz), 7.81 (1H, d, H-6)

Furthermore, an illustrative experiment of the preparation of acaprazol-1″′-ester-3″′-ester derivative of the formula (VII) accordingto the ninth aspect of this invention is explained with reference to thefollowing Example 11.

Example 11 (a) Synthesis of5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-methyl-esterderivative (the compound corresponding to the derivative of the formula(XXI) where R⁷ is methyl group)

The 5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol [the compound ofthe formula (XIV)] (60.7 mg) as obtained in Example 9(a) was dissolvedin N,N-dimethylformamide (1.8 ml). To the solution obtained were addedtriethylamine (0.034 ml) and as the esterifying reagent, methanol(0.0065 ml) and N,N-bis(2-oxo-3-oxazolidinyl)phosphinic chloride (31.5mg), and the resulting mixture was subjected to the reaction intended atroom temperature for 2 hours (for the methyl-esterification reaction).

The resulting reaction solution was concentrated to dryness and theresidue was extracted with chloroform. The chloroform extract was washedwith water, dried over anhydrous sodium sulfate and concentrated todryness. The residue obtained was purified by a silica-gel columnchromatography (developing solvent system: chloroform-methanol; 50:1).Thus, there was afforded the titled compound represented by thefollowing formula (XXI-1).

The yield was 30.9 mg (% yield; 50%).

[α]_(D) ¹⁹−33° (c 0.3, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol):

δ 1.27, 1.37, 1.42, 1.56 (each 3H, s, methyl in the isopropylidenegroup), 1.43 (9H, s, methyl in the t-butoxycarbonyl group), 2.50 (3H, s,NMe-5″′), 3.15 (3H, s, NMe-8″′), 3.36 (3H, s, COOMe), 5.27 (1H, s,H-1″), 5.71 (1H, d, H-5, J_(5,6)=8 Hz), 5.86 (1H, s, H-1′), 7.68 (1H, d,H-6).

(b) Synthesis of5″-N-Boc-2′,3′;2″,3″-di-O-isopropylidene-caprazol-1″′-methyl-ester-3″′-dodecanoyl-esterderivative (the compound included within the derivative of the formula(XXII) above)

The compound of the formula (XXI-1) (30.7 mg) as obtained in the step(a) above was dissolved in dichloromethane (0.56 ml), and to theresultant solution, under ice-cooling, were added4-dimethylaminopyridine (10.1 mg) and as the acylating reagent,dodecanoyl chloride (0.014 ml). The mixture so obtained was subjected tothe reaction intended under ice-cooling. After the reaction of 5 hours,there were further added 4-dimethylaminopyridine (6.5 mg) and dodecanoylchloride (0.0094 ml), and the acylation reaction was proceeded further.

After the reaction of 7 hours, methanol (0.02 ml) was added to thereaction solution, and then the resulting solution was diluted withchloroform. The so diluted solution was washed with 10% aqueouspotassium hydrogen sulfate solution and then with water, and dried overanhydrous sodium sulfate and then concentrated to dryness. The residueobtained was purified by a silica-gel column chromatography (developingsolvent system:chloroform-methanol, 100:0→100:1). Thus, there wasafforded the titled compound (26.5 mg; yield: 70%).

(c) Synthesis of caprazol-1″′-methyl-ester-3″′-dodecanoyl-esterderivative (corresponding to Compound VII-G in Table 11) of thefollowing formula (VII-2)

The compound obtained in the step(b) above (26.5 mg) was dissolved in amixture of trifluoroacetic acid-methanol (8:2) (0.25 ml), and theresulting solution was subjected to the reaction intended at roomtemperature for 4 hours (for the deprotection reaction). The resultingreaction solution was concentrated to dryness, and to the resultantresidue was added diethyl ether, and the insolubles formed were washedwith diethyl ether. Thus, there was yielded the titled compound in theform of an addition salt of bis-trifluoroacetic acid (23.8 mg; yieldfrom the compound of the formula (XXI-1) of the step (a) above; 60%).

[α]_(D) ²⁰+6° (c 1, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol):

δ 0.90 (6H, t, N(CH₂)₁₁ Me and (CH₂)₁₀ Me, J=7 Hz), 2.38 (2H, t, CH₂(CH₂)₉Me, J=7 Hz), 2.46 (3H, s, NMe-5″′), 3.13 (3H, s, NMe-8″′), 3.68(3H, s, COOMe), 5.20 (1H, s, H-1″), 5.42 (1H, br. s, H-3″′), 5.71 (1H,d, H-1′, J_(1′,2′)=2 Hz), 5.75 (1H, d, H-5, J_(5,6)=8 Hz), 7.71 (1H, d,H-6).

Further, there are given in the following ¹H-NMR spectrum data (indeutero-dimethylsulfoxide, TMS internal standard) of Compound VII-A,Compound VII-C, Compound VII-E and Compound VII-R which are includedwithin the general formula (VII) above and shown in Table 11 above asconcrete examples of these compounds.

Compound VII-A

δ 0.86 (3H, t, CH₃ (CH₂)₅CO, J=7 Hz), 2.29 (3H, s, NMe-5″′), 3.02 (3H,s, NMe-8″′), 5.02 (1H, s, H-1″), 5.52 (1H, s, H-1′), 5.66 (1H, d, H-5,J=8 Hz), 7.72 (1H, d, H-6, J=8 Hz), 11.33 (1H, s, NH-3).

Compound VII-C

δ 0.86 (3H, t, CH₃ (CH₂)₇CO, J=7 Hz), 2.29 (3H, s, NMe-5′″), 3.02 (3H,s, NMe-8″′), 5.02 (1H, s, H-1″), 5.52 (1H, d, H-1′, J=˜2 Hz), 5.66 (1H,dd, H-5, J=2, 8 Hz), 7.72 (1H, d, H-6, J=8 Hz), 11.33 (1H, d, NH-3, J=2Hz).

Compound VII-E

δ 0.86 (3H, t, CH₃ (CH₂)₉CO, J=7 Hz), 2.28 (3H, s, NMe-5′″), 3.01 (3H,s, NMe-8″′), 5.02 (1H, s, H-1″), 5.52 (1H, s, H-1′), 5.66 (1H, d, H-5,J=8 Hz), 7.73 (1H, d, H-6, J=8 Hz), 11.33 (1H, s, NH-3).

Compound VII-R

δ 0.85 (3H, t, CH₃ (CH₂)₇CH═, J=7 Hz), 2.28 (3H, s, NMe-5′″), 3.01 (3H,s, NMe-8″′), 5.02 (1H, s, H-1″), 5.52 (1H, s, H-1′), 5.66 (1H, d, H-5,J=8 Hz), 7.72 (1H, d, H-6, J=8 Hz), 11.33 (1H, s, NH-3).

Next, there are given an experimental example of the preparation of auridine derivative of the formula (IX) above from caprazol according tothe eleventh aspect of this invention as well as an experimental exampleof the preparation of an imidazolidinone derivative of the formula(VIII) above from the 5″-N-Boc-protected derivative of the said uridinederivative according to the tenth aspect of this invention withreference to Example 12 given below.

Example 12 (a) Preparation of the Uridine Derivative of the FollowingFormula (IX)

Caprazol (100.9 mg) was dissolved in 40% aqueous methylamine solution(3.0 ml) and the resulting mixture was subjected to the reactionintended at room temperature for 19 hours. The resulting reactionsolution was concentrated under a reduced pressure and then dried undera reduced pressure. The resulting residue was washed with a mixedsolvent of chloroform-diethyl ether (1:1). The residue thus washed wasdried under a reduced pressure to give a solid (93.8 mg). The solid waschromatographed for the purification by passing through a column packedwith Amberlite (Registered Trade Mark) CG-50 (NH₄ ⁺ form), followed bydeveloping the column with water. The resulting eluate fractionscontaining the desired compound are concentrated under a reducedpressure and then the concentrated was dried under a reduced pressure,thus affording the titled compound (72.5 mg; yield: 68%).

[α]_(D) ²⁰+30° (c 1, H₂O)

¹H-NMR spectrum (500 MHz, in D₂O)

δ 2.30 (3H, s, NMe-6′), 2.50 (3H, s, NMe-2″′), 2.61 (3H, s, NMe-7′),3.39 (1H, d, H-2″′, J_(2″′,3″′)=4 Hz), 3.47 (1H, d, H-6′, J_(5′,6′)=9Hz), 5.06 (1H, d, H-1″, J_(1″,2″)=2 Hz), 5.54 (1H, d, H-1′,J_(1′,2′)=2.5 Hz), 5.67 (1H, d, H-5, J_(5,6)=8 Hz), 7.61 (1H, d, H-6).

(b) Synthesis of 5″-N-Boc-uridine derivative represented by thefollowing formula (IXa)

5″-N-Boc-caprazol (14.6 mg) as obtained in Example 8(a) was dissolved inaqueous 40% methylamine solution (0.73 ml) and the resulting mixture wassubjected to the reaction intended at room temperature for 2 days. Theresulting reaction solution was concentrated under a reduced pressureand then dried under a reduced pressure, thus affording the titledcompound (13.7 mg; yield: 90%).

[α]_(D) ²¹+13° (c 1.5, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol)

δ 1.49 (9H, s, methyl in t-butoxycarbonyl group), 2.46 (3H, s, NMe-6′),2.73 (3H, s, NMe-7′), 2.75 (3H, s, NMe-2″′), 5.09 (1H, br. s, H-1″),5.77 (1H, d, H-5, J_(5,6)=8 Hz), 5.88 (1H, d, H-1′, J_(1′,2′)=4 Hz),7.95 (1H, d, H-6).

(c) Synthesis of N-protected-imidazolidinone derivative represented bythe following formula (VIIIa-1) (corresponding to a compound includedwithin the derivative of the formula (VIIIa) above)

The N-protected-uridine derivative of the formula (IXa) as obtained inthe step (b) above (167.6 mg) was dissolved in N,N-dimethylformamide(2.5 ml), and to the resultant solution was added dodecyl isocyanate(0.63 ml) as an alkyl isocyanate of the formula (XXIV) above. Theresulting mixture was subjected to the reaction intended at roomtemperature. After 1 hour from the start of the reaction, a furtheramount (0.63 ml) of dodecyl isocyanate was added and the reaction wasproceeded for further 3 hours. The insolubles so deposited were filteredoff and the remaining reaction solution was concentrated under a reducedpressure. The resulting concentrate was extracted with chloroform, andthe chloroform extract was washed with an aqueous saturated sodiumsulfate solution, dried over anhydrous sodium sulfate and thenconcentrated under a reduced pressure. The resulting residue was washedwith hexane and dried under a reduced pressure, to give a solid (261mg). This solid was purified by a silica-gel column chromatography(developing system:chloroform-methanol-water, 9:1:0.1), thus affordingthe titled compound of the formula (VIIIa-1) (32.5 mg) (yield: 15%).

¹H-NMR spectrum (500 MHz, in deutero-methanol)

δ 0.89 (3H, t, N(CH₂)₁₁ Me, J=7 Hz), 2.47 (3H, s, NMe-6′), 2.75 (3H, s,NMe-7′), 2.99 (3H, s, NMe-2″′), 5.09 (1H, d, H-1″).

(d) Synthesis of the Imidazolidinone Derivative of the Following Formula(VIII-1) (Corresponding to Compound VIII-G Shown in Table 13 Above)

The N-protected imidazolidinone derivative of the formula (VIIIa-1)(32.5 mg) obtained in the step (c) above was dissolved in a mixedsolution of trifluoroacetic acid-methanol (8:2) (0.33 ml). The resultingmixture was subjected to the reaction intended at room temperature for 2hours (for the deprotection reaction). The resulting reaction solutionwas concentrated under a reduced pressure and the resulting residue waswashed with diethyl ether and then dried under a reduced pressure, thusaffording the titled compound of the formula (VIII-1) in the form of anaddition salt of bis-trifluoroacetic acid (32.5 mg; yield: 88%).

[α]_(D) ²⁰+13° (c 2, methanol)

¹H-NMR spectrum (500 MHz, in deutero-methanol)

δ 0.89 (3H, t, N(CH₂)₁₁ Me, J=7 Hz), 5.16 (1H, d, H-1″), 7.74 (1H, d,H-6, J_(5,6)=8 Hz).

Furthermore, the preparation of antibiotics, caprazamycins A-F which areused as starting materials in the preparation of caprazene and caprazolis now illustrated with reference to the following Reference Example 1.

Reference Example 1 Preparation of Antibiotics, Caprazamycins A-F

Streptomyces sp. MK730-62F2 (deposited under the depository number ofFERM BP-7218), which had been cultured on an agar slant culture medium,was inoculated in a culture medium which had been prepared by placinginto Erlenmeyer flasks (500 ml-capacity) 110 ml portions of a liquidculture medium comprising 2% galactose, 2% dextrin, 1% glycerine, 1%Bacto-Soyton (a product of Difco Co.), 0.5% corn steep liquor, 0.2%ammonium sulfate and 0.2% calcium carbonate (adjusted to a pH of 7.4)and sterilizing the culture medium in the flasks at 120° C. for 20minutes in a usual manner, before the inoculation of the strainMK730-62F2 was done. The liquid culture medium so inoculated was thensubjected to shaking cultivation with rotation at 30° C. for 2 days,whereby giving a seed culture broth as intended.

In a tank fermenter (30 L-capacity), there was prepared a culture medium(15 L) comprising 2.4% tomato paste (a product of Kagome Co.), 2.4%dextrin, 1.2% yeast extract (a product of Oriental Co.) and 0.0006%cobalt chloride (adjusted to a pH of 7.4), which was then sterilized togive a productive culture medium. To this productive culture medium wasinoculated a 2% proportion of the above-mentioned seed culture broth.The cultivation of the said strain was conducted in the tank fermenterat 27° C., with aeration of 15 L of air per minute and stirring speed of200 rpm for 6 days.

The resulting culture broth was centrifuged to separate the culturebroth filtrate (12 L) from the cultured microbial cells. Subsequently,methanol (6 L) was added to the microbial cells and the resultingmixture was well stirred to extract from the microbial cells the alreadyknown antibiotics, caprazamycins A, B, C, E and F, and the novelantibiotics, caprazamycins D, G, D1 and G1 into methanol (hereinafter,sometimes, these antibiotics, caprazamycins A, B, C, D, E, F, G, D1 andG1, are generically described as caprazamycins).

The culture broth filtrate and the methanolic extract of the cellsobtained as above were combined together, and the resulting mixture (18L) was passed through a column packed with 750 ml of a syntheticadsorbent resin made of aromatic polymer, namely Diaion HP-20 (a productof Mitsubishi Chemical Co., Japan) to adsorb caprazamycins therein.

Through this Diaion HP-20 column containing the caprazamycins soadsorbed, were passed 2.25 L each of deionized water, 50% aqueousmethanol, 80% aqueous methanol, 80% aqueous acetone and acetone, inorder. The caprazamycins were eluted from the column, mainly in theeluate fractions as eluted with 80% aqueous acetone. In addition, theeluate fractions as eluted with the 50% aqueous methanol and with the80% aqueous methanol also contained caprazamycins. These eluatefractions as eluted with the two aqueous methanolic solvents werecombined together and the mixture was again passed through a column ofDiaion HP-20 (750 ml), whereby caprazamycins were adsorbed in theadsorbent of this column. Then, elution of this column was effected bypassing 80% aqueous methanol (2.25 L) therethrough. Subsequently, thecolumn was eluted with 80% aqueous acetone (2.25 L). The resultingeluate as eluted with 80% aqueous acetone was combined with the firsteluate as eluted with 80% aqueous acetone in the first stage column andthe resulting mixture was concentrated to dryness under a reducedpressure, thus affording a partially purified product comprisingcaprazamycins (10.1 g).

The partially purified product (10.1 g) containing caprazamycins wasdissolved in a mixed solvent (50 ml) of chloroform-methanol (1:2), towhich solution was added Kieselgur (Art. 10601, a product of Merck &Co.) (50 ml) and the solvent was removed by concentration to drynessunder a reduced pressure. The resulting solid residue obtained byadsorbing the caprazamycins in the Kieselgur was placed on the top of asilica-gel column (54 mm inner diameter and 200 mm long) to be subjectedto a chromatography. The development treatment was made, in order, withchloroform-methanol-water (4:1:0.1), chloroform-methanol-water (2:1:0.2)and chloroform-methanol-water (1:1:0.2) (1.35 L in each time).

The eluates from the silica-gel column were collected each in fractionsby means of fraction collector, so that fractions Nos. 1 to 53 werecollected each in 20 g portions and fractions Nos. 54 to 117 werecollected each in 19 g portions. In this way, the active fractionscontaining caprazamycins A, B, C, D, E, F and G were eluted in fractionsNos. 66 to 83 and the active fractions containing caprazamycins D1 andG1 were eluted in fractions Nos. 84 to 144. The fractions Nos. 66 to 83containing caprazamycins A, B, C, D, E, F and G were combined togetherand concentrated to dryness under a reduced pressure to yield apartially purified product comprising caprazamycins A, B, C, D, E, F andG (625.3 mg). The fractions Nos. 54 to 117 were also combined togetherand concentrated to dryness under a reduced pressure to yield apartially purified product comprising caprazamycins D1 and G1 (1.28 g).

Subsequently, the partially purified product containing caprazamycins A,B, C, D, E, F and G was subjected to treatments for the isolation andpurification of the respective compounds from one another. Thus,methanol (5 ml) was added to the said partially purified product above(625.3 mg) and the resulting solution was allowed to stand at 5° C.under cold and dark conditions, whereby a fraction of precipitate asdeposited (537.3 mg) was obtained as a product which containscaprazamycins A, B, C, D, E, F and G. Then, the fraction of thedeposited precipitate containing caprazamycins A, B, C, D, E, F and Gwas purified by subjecting it to HPLC (CAPCELLPAK C18, φ20×250 mm, aproduct of Shiseido, Co.). In this HPLC, the development was effected by50% aqueous acetonitrile-0.05% formic acid as the development solvent(at a flow rate of 12.0 ml/min), whereby caprazamycin A was eluted after61-68 minutes, caprazamycin B was eluted after 52-60 minutes,caprazamycin C was eluted after 39-41 minutes, a mixture of caprazamycinD and caprazamycin G was eluted after 30-38 minutes, caprazamycin E waseluted after 25-28 minutes, and caprazamycin F was eluted after 22-25minutes of the development. The respective active fractions werecollected, separately and concentrated to dryness under a reducedpressure, thus to afford caprazamycin A (56.9 mg), caprazamycin B (90.3mg), caprazamycin C (19.7 mg), a mixture comprising caprazamycin D andcaprazamycin G (162.9 mg), caprazamycin E (30.3 mg) and caprazamycin F(25.5 mg), respectively.

Further, the mixture comprising caprazamycin D and caprazamycin Gobtained as above (162.9 mg) was purified by HPLC (CAPCELLPAK C18,φ20×250 mm, a product of Shiseido, Co.). In this HPLC, the developmentwas effected with the solvent system of 50% aqueous acetonitrile-0.025%trifluoroacetic acid (at a flow rate of 9.0 ml/min), wherebycaprazamycin D was eluted after 55-69 minutes and caprazamycin G waseluted after 48-53 minutes of the development. The respective activefractions were collected separately and then concentrated to drynessunder a reduced pressure, thus to afford caprazamycin D (69.7 mg) andcaprazamycin G (39.0 mg), respectively.

Further, the partially purified product (1.28 g) containingcaprazamycins D1 and G1 obtained as above was subjected to thetreatments for the isolation and purification of the respectivecompounds from each other by HPLC (CAPCELLPAK C18, φ20×250 mm, a productof Shiseido, Co.). In this HPLC, the development was effected with thesolvent system of 45% aqueous acetonitrile-0.05% trifluoroacetic acid(at a flow rate of 12.0 ml/min), whereby caprazamycins G1 and D1 wereeluted after 36-49 minutes of the development. These eluate fractionswere collected and concentrated to dryness under a reduced pressure,thus to afford a mixture of caprazamycin D1 and caprazamycin G1 (187mg). The said mixture was further subjected to HPLC (CAPCELLPAK C18,φ20×250 mm, a product of Shiseido, Co.) wherein the development waseffected with the solvent system of 44% aqueous acetonitrile-0.025%trifluoroacetic acid (at a flow rate of 9.0 ml/min), wherebycaprazamycin D1 was eluted after 46-52 minutes and caprazamycin G1 waseluted after 41-44 minutes of the development. These eluate fractionswere collected and concentrated to dryness under a reduced pressure,respectively, thus to afford each of caprazamycin D1 (54.1 mg) andcaprazamycin G1 (57.6 mg).

INDUSTRIAL APPLICABILITY

As described hereinbefore, caprazene and caprazol are now produced byhydrolysis of caprazamycins according to this invention. Caprazene andcaprazol are useful compounds for the production of semi-synthesizedderivatives having excellent antibacterial activities. According to thisinvention, further, there are synthesized novel compounds which are acaprazene-1″′-amide derivative of the formula (II), acaprazene-1″′-ester derivative of the formula (III), acaprazol-1″′-amide derivative of the formula (V), acaprazol-1″′-amide-3″′-ester derivative of the formula (VI), acaprazol-3″′-ester derivative and a caprazol-1″′-ester-3″′-esterderivative of the formula (VII) or an imidazolidinone derivative CP-IMof the formula (VIII). These derivatives of caprazene and caprazol haveexcellent antibacterial activities against a variety of bacteria and areuseful as antibacterial agents. Further, the uridine derivative of theformula (IX) as obtained according to this invention is useful as novelintermediate compound utilizable in the syntheses of a variety of novelcompounds.

1. Caprazene which is the compound represented by the following formula(Ia):

wherein Me stands for methyl group, and A is a hydrogen atom, andwherein said compound has the ¹H-NMR and ¹³C-NMR data as set forth inTable 15.